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Title: TOS Standard Operating Procedure: APB - Measurement of Aboveground Productivity

Date: 03/17/2021
for Agricultural Crops

NEON Doc. #: NEON.DOC.001714 Author: C. Meier Revision: E

TOS STANDARD OPERATING PROCEDURE: APB -


MEASUREMENT OF ABOVEGROUND PRODUCTIVITY FOR
AGRICULTURAL CROPS

PREPARED BY ORGANIZATION DATE


Dave Barnett SCI 02/07/2021
Courtney Meier SCI 01/03/2019
Katie Jones SCI 03/01/2016

APPROVALS ORGANIZATION APPROVAL DATE


Kate Thibault SCI 03/16/2021

RELEASED BY ORGANIZATION RELEASE DATE


Frank Mocilac CM 03/17/2021

See configuration management system for approval history.

The National Ecological Observatory Network is a project solely funded by the National Science Foundation and managed under cooperative agreement by Battelle.
Any opinions, findings, and conclusions or recommendations expressed in this material are those of the author(s) and do not necessarily reflect the views of the
National Science Foundation.

Template_NEON.DOC.002626 Rev K
Title: TOS Standard Operating Procedure: APB - Measurement of Aboveground Productivity
Date: 03/17/2021
for Agricultural Crops

NEON Doc. #: NEON.DOC.001714 Author: C. Meier Revision: E

Change Record

REVISION DATE ECO # DESCRIPTION OF CHANGE


A 09/15/2016 ECO-04141 Initial release
• Section 4, Timing: Clarified that 2+ harvests per plot
are required if plot is planted with multiple crops per
season, or is fallow for part of year and then planted.
• Section 6, Equipment: Added folding ruler, loppers,
and spring scales.
B 07/12/2017 ECO-04624
• SOP B: Weighing of total fresh mass, creation of sub-
sample, and weighing of subsample fresh mass now
carried out in the field with spring scales, rather than
lab, and can be employed for all crop-types. Prevents
transport of large volumes of clipped mass.
• Section 4.1, Sampling Frequency and Timing: Re-
organized and added more examples to illustrate
desired plot numbers based on learned experience at
various sites.
• Section 4.2, Onset and Sessation of Sampling: Added
guidance for crops that do not appear in Table 2, and
for which USDA timing data are not available (e.g.,
millet in D02).
• Section 6.4, Estimated Time: Added updated
C 02/16/2018 ECO-05405 standardized time estimates per SOP.
• SOP B: Updated text to reflect mobile app-based
workflow and barcodes
• SOP D: Added optional workflow for using barcodes
with samples
• SOP D.1, Drying and Weighing: Clarified that
ovenStartTime and ovenEndTime are only recorded for
initial drying period.
• Appendix A.1: Added LAJA specific guidance for
sunflower crop.

Template_NEON.DOC.002626 Rev K
Title: TOS Standard Operating Procedure: APB - Measurement of Aboveground Productivity
Date: 03/17/2021
for Agricultural Crops

NEON Doc. #: NEON.DOC.001714 Author: C. Meier Revision: E

• Added SOP A.2.1: Integration of Plant Belowground


Biomass and Clip-Harvest at agricultural sites.
• SOP B: Sampling areas for synchronized Plant
Belowground Biomass sampling are rotated along
with the crop clip-strip. SOP now matches training
materials.
• SOP B.1: Subsampling procedure for high-volume
D 01/29/2019 ECO-06002
crops modified to reflect easier workflow developed
in the field. Illustrations of workflow added.
• SOP B.2: Added guidance when rows cannot be
discerned for a portion of the plot.
• Added SOP C.1 to specify Field record finalizing and
auto-creation of lab records occurs after sort-
checking
• Updated to new template (NEON.DOC.002626vK).
E 03/17/2021 ECO-06539 • SOP B.1: Changed boutNumber to weekBoutBegan to
simplify bout naming convention.

Template_NEON.DOC.002626 Rev K
Title: TOS Standard Operating Procedure: APB - Measurement of Aboveground Productivity
Date: 03/17/2021
for Agricultural Crops

NEON Doc. #: NEON.DOC.001714 Author: C. Meier Revision: E

TABLE OF CONTENTS
1 DESCRIPTION ................................................................................................................................ 1
1.1 Overview ................................................................................................................................... 1
1.2 Purpose ..................................................................................................................................... 1
1.3 Scope ......................................................................................................................................... 1
1.4 Applies To .................................................................................................................................. 2
1.5 Acknowledgments ..................................................................................................................... 2
2 RELATED DOCUMENTS AND ACRONYMS ...................................................................................... 2
2.1 Applicable Documents .............................................................................................................. 2
2.2 Reference Documents ............................................................................................................... 2
2.3 Acronyms .................................................................................................................................. 3
2.4 Definitions ................................................................................................................................. 3
3 METHOD ...................................................................................................................................... 4
4 SAMPLING SCHEDULE ................................................................................................................... 4
4.1 Sampling Frequency and Timing ............................................................................................... 4
4.2 Criteria for Determining Onset and Cessation of Sampling ...................................................... 6
4.3 Timing for Laboratory Processing and Analysis ........................................................................ 8
5 SAFETY ......................................................................................................................................... 9
6 PERSONNEL ................................................................................................................................ 11
6.1 Training Requirements ............................................................................................................ 11
6.2 Specialized Skills ...................................................................................................................... 11
7 STANDARD OPERATING PROCEDURES ........................................................................................ 12
SOP A PREPARING FOR SAMPLING ............................................................................................ 13
SOP B FIELD SAMPLING ABOVEGROUND BIOMASS OF AGRICULTURAL CROPS .......................... 17
SOP C POST-FIELD SAMPLING TASKS ......................................................................................... 29
SOP D LABORATORY PROCESSING OF AGRICULTURAL BIOMASS SAMPLES................................. 31
SOP E DATA ENTRY AND VERIFICATION ..................................................................................... 35
APPENDIX A EQUIPMENT ............................................................................................................. 36
APPENDIX B SITE-SPECIFIC CROP SAMPLING ................................................................................ 41

i
Title: TOS Standard Operating Procedure: APB - Measurement of Aboveground Productivity
Date: 03/17/2021
for Agricultural Crops

NEON Doc. #: NEON.DOC.001714 Author: C. Meier Revision: E

LIST OF TABLES AND FIGURES

Table 1. Summary of clip harvest sampling frequency and timing guidelines by plot type for agricultural
sites. .............................................................................................................................................................. 5
Table 2. Anticipated earliest harvest dates for unique site by crop combinations likely at NEON
agricultural sites. ........................................................................................................................................... 7
Table 3. Clip Strip dimension required for crops expected at NEON agricultural sites. Row spacing is for
typical plantings, and may vary from listed value. ..................................................................................... 20
Table 4. Equipment list – Equipment needed to prepare for sampling. .................................................... 36
Table 5. Equipment needed for a 2-person team to perform agricultural clip harvest sampling at one
plot. ............................................................................................................................................................. 37
Table 6. Equipment needed for post-field sampling tasks. ........................................................................ 39
Table 7. Equipment needed for processing agricultural biomass clip harvest samples in the laboratory. 40

Figure 1. Base of corn stalk, including belowground roots and aboveground brace roots; the latter are
also known as ‘prop’ roots. ........................................................................................................................... 3
Figure 2. Overview of the SOPs needed to collect agricultural biomass and productivity data. ............... 12
Figure 3. Modified clip cell layout when integrating plant belowground biomass sampling and
herbaceous biomass clip harvest at agricultural sites ................................................................................ 15
Figure 4. Expanded workflow diagram for field sampling the aboveground biomass of agricultural crops
(SOP B). Diagram supports and does not replace protocol text; most common workflow is outlined...... 17
Figure 5. Clip Strip establishment in an agricultural row crop ................................................................... 20
Figure 6. Subsampling routine for high-volume crops with large seeds (e.g., corn). Letters (a-j)
correspond to protocol steps...................................................................................................................... 24
Figure 7. Subsampling routine for high-volume crops with small seeds (e.g., cereals). Letters (a-f)
correspond to protocol steps...................................................................................................................... 26
Figure 8. Rough percent cover map of plot planted in both corn (dominant crop) and soybeans
(subdominant crop) .................................................................................................................................... 27
Figure 9. Expanded workflow diagram for sort-checking field-collected agricultural biomass prior to
oven-drying in the laboratory (SOP C) ........................................................................................................ 29
Figure 10. Expanded workflow diagram for drying and weighing clipped, sort-checked agricultural
biomass (SOP D) .......................................................................................................................................... 32

ii
Title: TOS Standard Operating Procedure: APB - Measurement of Aboveground Productivity
Date: 03/17/2021
for Agricultural Crops

NEON Doc. #: NEON.DOC.001714 Author: C. Meier Revision: E

1 DESCRIPTION

1.1 Overview

The Standard Operating Procedure (SOP) described in this document is an extension of the TOS Protocol
and Procedure: Measurement of Herbaceous Biomass (RD[04]). For crop clip-harvesting in the field
described here, the dimensions and orientation of typical Clip Strips used for ‘wild-type’ herbaceous
biomass clipping are modified. In contrast to Clip Strips for ‘wild-type’ vegetation, crop Clip Strips adopt
wider dimensions, and are oriented perpendicular to crop rows in order to account for the spatial
structure introduced by crop row-planting. The field procedure for crops is also modified from RD[04] to
include a subsampling routine. Because more area is sampled in the field, and crops can produce large
amounts of biomass, the subsampling routine allows smaller amounts of biomass per sample to be
transported back to the laboratory and placed into the drying ovens. Subsampling requires that
additional fresh weight data are collected in the field so that subsample dry weights can be scaled back
up to the entire sample collected in the field.

1.2 Purpose

This document outlines the procedure for measuring aboveground biomass in plots that have been
planted with:
• Corn
• Soybean
• Sorghum, or
• Cereal crops (Wheat, barley, millet, triticale, rye, oat, etc.)

Other crops, such as squash, tomatoes, peppers, etc., are outside the scope of this SOP, and must be
addressed on a case by case basis via NEON’s help ticket software.

1.3 Scope

This document provides a change-controlled version of an Observatory procedure. Documentation of


content changes (i.e. changes in particular tasks or safety practices) will occur via this change-controlled
document, not through field manuals or training materials.

Page 1
Title: TOS Standard Operating Procedure: APB - Measurement of Aboveground Productivity
Date: 03/17/2021
for Agricultural Crops

NEON Doc. #: NEON.DOC.001714 Author: C. Meier Revision: E

1.4 Applies To

The procedure described in this document is used in the following protocols:

Doc # Title
NEON.DOC.014037 TOS Protocol and Procedure: Measurement of Herbaceous Biomass

1.5 Acknowledgments

Quadrat dimensions for corn, soybean, and graminoid crops were taken from the Kellogg Biological
Station LTER protocol for Aboveground Net Primary Productivity. Many thanks to: Greg Chapman for
refining and streamlining the field subsampling routine in SOP B.

2 RELATED DOCUMENTS AND ACRONYMS

2.1 Applicable Documents

Applicable documents contain higher-level information that is implemented in the current document.
Examples include designs, plans, or standards.

AD[01] NEON.DOC.004316 Operations Field Safety and Security Plan


AD[02] NEON.DOC.004300 EHSS Policy, Program and Management Plan
AD[03] NEON.DOC.050005 Field Operations Job Instruction Training Plan
AD[04] NEON.DOC.000914 TOS Science Design for Plant Biomass, Productivity, and Leaf Area
Index
AD[05] NEON.DOC.004104 NEON Science Data Quality Plan

2.2 Reference Documents

Reference documents contain information that supports or complements the current document.
Examples include related protocols, datasheets, or general-information references.

RD[01] NEON.DOC.000008 NEON Acronym List


RD[02] NEON.DOC.000243 NEON Glossary of Terms
RD[03] NEON.DOC.002652 NEON Data Products Catalog
RD[04] NEON.DOC.014037 TOS Protocol and Procedure: Measurement of Herbaceous Biomass
RD[05] NEON.DOC.001574 Datasheets for TOS Protocol and Procedure: Measurement of
Herbaceous Biomass
RD[06] NEON.DOC.001717 TOS Standard Operating Procedure: TruPulse Rangefinder Use and
Calibration
RD[07] NEON.DOC.001025 TOS Protocol and Procedure: Plot Establishment
RD[08] NEON.DOC.001271 AOS/TOS Protocol and Procedure: Data Management

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Title: TOS Standard Operating Procedure: APB - Measurement of Aboveground Productivity
Date: 03/17/2021
for Agricultural Crops

NEON Doc. #: NEON.DOC.001714 Author: C. Meier Revision: E

2.3 Acronyms

Acronym Definition
ddh Diameter at decimeter height
NPP Net Primary Productivity
ISO International Organization for Standardization

All other acronyms used in this document are defined in RD[01].

2.4 Definitions

Brace roots: (Also called prop roots) Aerial roots originating along a corn stalk, above the soil surface
(Figure 1).

Ear: Pollinated female corn inflorescence comprised of cob, kernel, husk leaves and silk (style).

Fulcrum: Software tool used to create NEON electronic data entry applications.

High-Volume Crop: Functionally, high-volume crops are those that, when sampled and brought back to
the laboratory without subsampling, would overwhelm available oven space. Typically, it is not possible
to process high-volume crops within the specified 5 d cold storage limit without subsampling. Because
yields and growth rates can vary tremendously from year to year due to external factors, a crop may be
high-volume one year, and low-volume the next (e.g., soy).

Low-Volume Crop: Functionally, samples from low-volume crops can be dried in their entirety without
overwhelming available oven space, and all samples can be processed within the 5 d cold-storage limit
without subsampling.

Pod: The elongated seed vessel of the soybean plant.

ServiceNow: Software tool used for problem/incident tracking and resolution.

Figure 1. Base of corn stalk, including belowground roots and aboveground brace roots; the latter are also known
as ‘prop’ roots.

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Title: TOS Standard Operating Procedure: APB - Measurement of Aboveground Productivity
Date: 03/17/2021
for Agricultural Crops

NEON Doc. #: NEON.DOC.001714 Author: C. Meier Revision: E

3 METHOD

A combination of Distributed, Gradient, and Tower Plots may be used for collecting biomass and
productivity data from agricultural crops. The timing of agricultural biomass sampling is managed on a
per crop basis within each site, and the clip harvest of similar plots within a given crop type is targeted
to peak crop biomass, and is constrained by crop senescence, anticipated harvest date, or both. In
addition, multiple clip harvests per season are required to capture the combination of fallow cover and
agricultural crop productivity if farmers employ multiple crop rotations within a growing season (see
Section 4 for more details).

In the field, the primary distinction between the procedure for agricultural crops described here,
compared to that presented in RD[04] for ‘wild’ herbaceous vegetation, is that crops require larger,
variable Clip Strip dimensions, and Clip Strips are oriented perpendicular to crop rows. In contrast, ‘wild’
herbaceous vegetation uses a Clip Strip with fixed dimensions that is always oriented North/South.
Variable Clip Strip dimensions are required because planting in rows introduces spatial structure to the
biomass that is not adequately captured with a narrow Clip Strip.

Because agricultural plots may also support other herbaceous plants besides the crop of interest
(subsequently referred to in this document as ‘non-crops’), clipped vegetation within these larger Clip
Strips must still be sorted to functional group as in RD[04] if non-crops are present. For example, if a Clip
Strip contains wheat, as well as other plants belonging to several of the herbGroups defined in RD[04],
clipped wheat biomass will be separated from non-crop biomass, and clipped non-crop biomass will be
further sorted to herbGroup (i.e., CSG, WSG, LFB, etc.).

Because agricultural crops may attain relatively large stature, the laboratory procedure in this SOP
differs from RD[04] by introducing a subsampling method prior to oven-drying clipped biomass of high-
volume crops. This subsampling step allows for more efficient drying and processing of large sample
volumes. Low-volume crops are processed identically to ‘wild’ herbaceous vegetation. In addition, data
collected via this SOP are entered via the same data ingest mechanism as RD[04].

4 SAMPLING SCHEDULE

4.1 Sampling Frequency and Timing

Agricultural sites often practice crop rotation, which results in NEON plots located in different parcels,
with each parcel supporting a crop-specific planting and harvest schedule that changes from year to
year. Each crop that matures within the given sampling year will need to be sampled for biomass in
order to estimate the productivity of the site, and multiple sampling bouts are therefore required to
capture the productivity of crops planted and harvested in different seasons. For these reasons, the
following actions are highly advantageous in agricultural systems:

• Communicate and collaborate with the site host and/or farmer to determine and coordinate
harvest dates and sampling activities,

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Title: TOS Standard Operating Procedure: APB - Measurement of Aboveground Productivity
Date: 03/17/2021
for Agricultural Crops

NEON Doc. #: NEON.DOC.001714 Author: C. Meier Revision: E

• Monitor the Phenocam to determine crop maturation rates and potential harvest dates, and

• Consult the Site Management app for historical crop-specific harvest dates.
Table 1 provides an overview of spatial and temporal sampling strategy for agricultural sites.

Fallow Fields:

• Plots left fallow or planted with cover crops will need to be sampled in order to adequately
capture site-level productivity (see examples below).

• Weeds or cover crops in fallow plots that do not have an obvious row structure should be
clipped with the standard 2.0m x 0.1m clip strip, oriented North/South.

• For plots that are clip harvested for late-season crops, it is not necessary to schedule an
additional bout for clipping weeds that may sprout late in the season.
Table 1. Summary of clip harvest sampling frequency and timing guidelines by plot type for agricultural sites.

# Events per Yearly


Plot Type Plot Number Sampling Year Interval Sampling Start Sampling Stop

n=10 (min)* Once per crop Annual Beginning of crop Within 14d of
Distributed cycle or fallow senescence sampling start
n=20 (max) period
n=10 per crop
Tower
type (max)

* If all Distributed Plots are planted in the same crop type, a minimum of n=10 plots may be sampled (i.e., the 10
lowest Morton Order plots). If fewer than n=10 plots exist for a given crop type, all plots may be sampled. In the
event that these rules result in > 20 plots that should be clipped, consult with Science.

Example Agricultural Clip Harvest Scenarios:


These examples convey the general concept that biomass produced by different crop and non-crop
plants at different times of year should be accounted for with appropriately scheduled clip-harvest
bouts. Not all possible scenarios are covered; if it is unclear how to schedule for your site, create an
incident ticket and discuss with Science.
Multiple Crop Types Maturing at Different Times. A north temperate site has 10 plots planted
with cover-crop, 10 plots planted with winter wheat, and 10 plots planted with corn. Plots
planted with winter wheat would be harvested during the first bout (likely late May or early
June, approximately ISO week 22); cover-crop and corn plots would be harvested during a
second bout (likely mid to late August approximately ISO week 34). Cover-crop plots may be
harvested on a different schedule than corn if the cover-crop reaches peak biomass later than
the corn harvest.

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Title: TOS Standard Operating Procedure: APB - Measurement of Aboveground Productivity
Date: 03/17/2021
for Agricultural Crops

NEON Doc. #: NEON.DOC.001714 Author: C. Meier Revision: E

Early-Season Fallow Followed by Crop. A plot (Distributed or Tower) is fallow for the spring (i.e.,
nothing planted) and has mixed non-crop plants growing opportunistically in it. The plot is then
plowed and planted with a crop that matures in late summer or early autumn. Plots such as this
require a first bout for harvesting non-crop biomass in the spring, and a second bout for
quantifying crop biomass in the autumn.

Early-Season Crop Followed by Fallow. A plot (Distributed or Tower) is planted with winter
wheat that is harvested early to mid-summer. The plot is then left fallow, and opportunistic non-
crop plants grow, or is planted with a cover crop, and these plants senesce in the autumn. Plots
such as this require a first bout for quantifying early-season crop biomass, and a second bout for
assessing peak non-crop biomass (scheduled when greenness begins to decrease – i.e., when
plants first begin to senesce).

Multiple Crop Types and Desired Sample Size. At a site with n=30 Tower Plots, not all plots are
planted in crops, and for plots with crops, plot numbers across crop types are not equal. For
example, assume 3 Tower Plots are planted with winter wheat, 14 Tower Plots are planted with
corn, and the remaining 13 Tower Plots are located in perennial pasture/hay vegetation. In this
scenario, one clip harvest bout would be scheduled early-season for all 3 winter wheat plots,
another late-season bout would be scheduled to clip n=10 corn plots (the 10 lowest Morton
Order plots), and a third bout would be scheduled according to the dates in RD[04] for the
perennial pasture/hay vegetation. Winter wheat plots left fallow after harvest may be clipped
again during the late-season corn bout if non-crop biomass is present at that time. A similar
strategy would be employed for Distributed Plots planted the same way.

4.2 Criteria for Determining Onset and Cessation of Sampling

Sampling onset: Crop biomass typically peaks some time before plants begin to senesce, and a fraction
of total peak biomass mass is lost as senescence proceeds. Peak greenness data provided in Appendix D
of RD[04] may guide scheduling of sampling onset, but because multiple rotations are often planted
within a site, MODIS data should be considered as a guide only. A more reliable indicator for sampling
onset is to monitor plants for the beginning of senescence, and begin clipping as soon as the first leaves
begin to dry or yellow. For corn and soybeans, drying and yellowing usually begins with the lower, older
leaves.

Sampling cessation: As indicated in Table 1, sampling should be completed within 2 weeks of sampling
onset. In the event of unforeseen delays, sampling must be completed before the farmer harvests the
crop; if the farmer removes crop biomass before NEON clip harvest is complete, estimates of
productivity for the site will be severely compromised. Table 2 gives the earliest dates that farmers
typically harvest specific crops within NEON sites (data provided by USDA).

Occasionally, farmers will plant specialty crops for which USDA has not compiled harvest date data –
e.g., millet planted at the D02 BLAN site. In this event, to complete NEON Ag Clip Harvest sampling

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Title: TOS Standard Operating Procedure: APB - Measurement of Aboveground Productivity
Date: 03/17/2021
for Agricultural Crops

NEON Doc. #: NEON.DOC.001714 Author: C. Meier Revision: E

activities before the farmer harvests the crop, opportunistically monitor crop maturation through time
while conducting other sampling, and/or maintain communication with the farmer about anticipated
harvest dates.

Table 2. Anticipated earliest harvest dates for unique site by crop combinations likely at NEON agricultural sites.
Data come from long-term USDA records aggregated at the state level. Sites with crops other than those listed
here are covered in the site-specific modification appendix at the end of this document (e.g., D04 LAJA).

Harvest most active


Domain Site(s) Crop Harvest begin date dates
Corn for grain 8/25 9/5 – 10/25
Corn for silage 8/5 8/30 – 10/1
BLAN
D02 Soybeans 10/5 10/25 – 11/25
Wheat, winter 6/5 6/20 – 7/15
SERC Soybeans 10/5 10/18 – 11/15
Corn for grain 7/25 8/15 – 9/5
D03 JERC Soybeans 10/1 11/1 – 11/25
Wheat, winter 5/20 6/1 – 6/15
Barley, fall 6/10 6/15 – 7/1
Barley, spring 6/20 6/25 – 7/1
Corn for grain 9/5 9/20 – 10/20
D06 KONA
Sorghum for grain 9/15 10/10 – 11/5
Soybeans 9/20 10/5 – 10/25
Wheat, winter 6/15 6/20 – 7/10
Barley, spring 7/30 8/8 – 8/23
Corn for silage 8/31 9/12 – 9/28
Corn for grain 9/29 10/10 – 10/27
D09 NOGP
Soybeans 9/16 9/26 – 10/11
Wheat, durum 8/9 8/21 – 9/9
Wheat, spring 8/4 8/14 – 9/1
Barley, spring 6/10 7/25 – 9/5
Corn for silage 9/1 9/10 – 9/30
Corn for grain 10/1 10/15 – 11/10
D10 STER
Sorghum for grain 9/25 10/10 – 11/15
Sorghum for silage 9/1 9/5 – 9/20
Wheat, winter 6/25 7/10 – 7/20

Dates listed in Table 2 may be used for hiring and high-level scheduling purposes if Field Operations
obtains information from the farmer with respect to which crops will be planted in the coming year.
However, with respect to scheduling the exact week in which clip harvests will take place for a particular
crop once the growing season is underway, bear in mind that weather can cause significant deviations
from the averages presented in Table 2.

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Title: TOS Standard Operating Procedure: APB - Measurement of Aboveground Productivity
Date: 03/17/2021
for Agricultural Crops

NEON Doc. #: NEON.DOC.001714 Author: C. Meier Revision: E

4.3 Timing for Laboratory Processing and Analysis

Because clipped biomass continues to be biologically active after clipping and before drying (i.e. plant
cells continue to respire and therefore lose mass), it is important to place clipped samples into the
drying oven as soon as possible after clipping occurs. For Herbaceous Biomass (RD[04]), clipped plants
are kept in cold storage after clipping and until samples can be placed in the drying oven. Similar to
RD[04], freshly clipped crop samples or subsamples must be placed in the drying oven within 5 d of
clipping in the field, and crop biomass must be kept in cold storage, same as in RD[04]. Once samples are
dry, timing considerations for weighing are identical to RD[04].

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Title: TOS Standard Operating Procedure: APB - Measurement of Aboveground Productivity
Date: 03/17/2021
for Agricultural Crops

NEON Doc. #: NEON.DOC.001714 Author: C. Meier Revision: E

5 SAFETY

This document identifies procedure-specific safety hazards and associated safety requirements. It does
not describe general safety practices or site-specific safety practices.

Personnel working at a NEON site must be compliant with safe field work practices as outlined in the
Operations Field Safety and Security Plan (AD[02]) and EHS Safety Policy and Program Manual (AD[01]).
Additional safety issues associated with this field procedure are outlined below. The Field Operations
Manager and the Lead Field Technician have primary authority to stop work activities based on unsafe
field conditions; however, all employees have the responsibility and right to stop their work in unsafe
conditions.

Agricultural Chemicals

Common agricultural chemicals include pesticides, insecticides, fungicides, and herbicides and residues
of these chemicals may be found on active agricultural sites. Their hazards may remain after fields have
been treated. These chemicals can be on plants, in soil, and sometimes in irrigation systems used to
apply hazardous chemicals. Chemical residue cannot always be seen, and may also be carried by the
wind.

It is important to determine if it is safe to enter a field where pesticides have been applied prior to
beginning work. Attempts should be made to determine pesticide application schedule and the
restricted entry interval (REI) of applied pesticides prior to entering agricultural fields or areas where
pesticide use is expected. When entering agricultural fields or other areas, check for posted warning
signs at access roads, field borders adjacent to public areas and established walking routes where
agricultural workers enter the area.

Pesticide Safety Training is required if entering/working in a field that has been sprayed within the
last 30 days. The following precautions must be followed when re-entering areas within 30 days after
expiration of restricted entry interval (REI):

• Wear full-length pants, long-sleeved shirts, a hat, socks, and non-leather gloves and work shoes.
Leather can absorb chemicals from plants and the soil.

• Wash gloves and work boots/shoes prior to removing, if possible.

• Wash work clothes separately from other laundry using hot water and laundry detergent.
Always wash clothes, boots, and gloves before wearing them again.

• Wash face, neck, hands, and arms as soon as possible, after potential exposure to agricultural
chemicals.
• NEVER enter a field that has been posted with a “No Re-Entry” warning sign! Keep in mind sign
verbiage may vary.

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Title: TOS Standard Operating Procedure: APB - Measurement of Aboveground Productivity
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for Agricultural Crops

NEON Doc. #: NEON.DOC.001714 Author: C. Meier Revision: E

• DO NOT smoke while working in a treated field, regardless of when the area was sprayed with
the hazardous chemical.

• DO NOT carry food of any kind into treated work area.

• If exposed or if exhibiting any signs or symptoms of exposure:


o Seek the Safety Data Sheet for the hazardous chemical that was used on the field. If
medical assistance isn’t immediately available, follow the first-aid directions on the label
or included with the Safety Data Sheet.
o Inform the Domain Manager of any suspected exposure. The DSF should have Safety
Data Sheets on hand for chemicals used in the fields in which NEON plots are located.
Laser Rangefinder

A laser rangefinder/hypsometer/compass instrument may be used to navigate to clip cells within plots.
Safety considerations for this instrument include:
• Avoid staring directly at the laser beam for prolonged periods. The rangefinder is classified as
eye-safe to Class 1 limits, which means that virtually no hazard is associated with directly
viewing the laser output under normal conditions. As with any laser device, however,
reasonable precautions should be taken in its operation. It is recommended that you avoid
staring into the transmit aperture while firing the laser.
• Never attempt to view the sun through the scope. Looking at the sun through the scope may
permanently damage the eyes.

Sharp Blades

Sharp-bladed pruners and/or loppers may be used to clip and subsample crop plants. Safety
considerations for these tools include:
• Select the correct tool for the job.
• Use personal protective equipment, to include gloves, safety glasses, and work boots to
minimize injuries in the field.
• Assure all personnel working in the area are aware of the use of the sharp tools, and keep all
sharp blades safely away from others.
• Maintain good posture and do not twist or stretch body awkwardly while making cuts with a
pruner or lopper.

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6 PERSONNEL

6.1 Training Requirements

All technicians must complete required safety training as defined in the NEON Training Plan (AD[04]).
Additionally, technicians must complete procedure-specific training for safety and implementation of
this procedure as required in Field Operations Job Instruction Training Plan (AD[05]).

Additional training is required for working in areas where agricultural chemicals have or may have been
applied.

For the field component of this protocol, technicians must be trained in navigating to points in the field
with a GPS and manual methods. Most critically, technicians must be trained to quickly identify common
herbaceous species that inhabit agricultural fields within the region. Because different herbaceous
functional groups can be sensitive indicators of ecosystem responses to global change (e.g. N deposition,
warming, rising CO2), it is very important that field technicians within a domain can accurately and
quickly identify C3 and C4 graminoids as well as identify leguminous and non-leguminous forbs within
that domain.

Training for both the field and laboratory work must emphasize the importance of consistent, detailed
labeling of all samples, including proper use of barcodes if barcodes are used. Improper or inconsistent
labeling and sample tracking is the most common and problematic error associated with this work!

6.2 Specialized Skills

The lead plant technician must possess the demonstrated ability to identify crops to species, and to
identify non-crop species inhabiting cropped plots to functional group – either via visual inspection, or
via visual inspection in combination with a dichotomous or polyclave key.

• Identification of all leguminous forbs to functional group, in the absence of flowers, is required.
• Identification to species is not required for non-leguminous forbs and woody stemmed plants.
• Identification to species is required for cool-season (C3) and warm-season (C4) graminoid
functional groups. Technicians should be able to identify graminoids vegetatively.
To identify non-crop species, ideally each team member should know how to use diagnostic traits and a
dichotomous or polyclave key.

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7 STANDARD OPERATING PROCEDURES

Standard operating procedures provide specific instruction and suggestions for the generation of data
that is aligned with the goals of this document (Figure 2).

Figure 2. Overview of the SOPs needed to collect agricultural biomass and productivity data.

SOP A

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SOP A Preparing for Sampling

A.1 Sampling Equipment Preparation and Checklist

• See RD[04] for equipment preparation.

• Use of the laser rangefinder is only necessary at agricultural sites if plot markers are not present,
and full plot delineation is required.

A.2 Early-season Preparation at Agricultural Sites

• Check with the Domain Manager to ensure that the farmer (or site host) has approved
implementation of this SOP within target plots.

• If density of mature vegetation precludes easy navigation and Clip Strip delineation (e.g., in
mature corn crops), marking of plot corners and delineation of Clip Strips may occur earlier in
the season while plants are young and line-of-sight is not obscured.
o Temporarily record GPS coordinates for locations marked in this manner to enable re-
location during sampling.

SOP A

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SOP A.2.1 Integrating Plant Belowground Biomass and Clip-Harvest at Agricultural Sites

Delineation and flagging of sampling areas for both Plant Belowground Biomass Sampling and the
Agricultural Biomass SOP should be carried out at the same time regardless of which protocol is
executed first.

1. Bring a 3 m long folding ruler, or equivalent rigid measuring device, and 0.5m x 0.5m frames
used to lay out the belowground biomass sampling areas.
2. Locate the SW corner of the clip cell. When integrating Plant Belowground Biomass and clip-
harvest at agricultural sites, the Clip List coordinate will serve as the SW corner of the clip cell
rather than the clip strip. For example, the left-most flag in Figure 3 is placed at the coordinate
provided in the Clip List.
a. Clip List coordinates at Agricultural sites have the standard meaning when Plant
Belowground Biomass sampling is NOT integrated with the Ag clip-harvest.
3. Rotate clockwise until you are facing perpendicular to crop rows (Figure 3).
4. Use the rigid measuring stick to lay out the 3 m long left side of a 3.0m x 0.5m clip cell.
5. Use the 0.5m x 0.5m frames to layout the plant belowground biomass sampling areas at either
end of the clip cell. Flag the lower-left corner of the cell and the upper-right corner of the cell.
a. Flagging should remain if soil sampling occurs prior to agricultural clip harvest and
allowed by the site host.
b. Note that the clip-strip may be wider than 0.5 m for some crop types.
6. Delineate a clip strip of the appropriate dimensions; the long edge of the clip strip should
remain perpendicular to crop rows.

SOP A

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Figure 3. Modified clip cell layout when integrating plant belowground biomass sampling and herbaceous biomass
clip harvest at agricultural sites. Orange dashes indicate the rotated clip cell. The red flag on the left is placed at
the coordinate provided in the Clip List.

SOP A

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A.3 Sample Labels and Identifiers

By default, this procedure considers each clip cell harvested on a unique date to be a sample, and each
herbGroup/crop is a subsample. Subsamples are labeled with the location, date, and herbGroup of the
collected subsample. In addition to labeling the subsample with human readable information, each
subsample may also be associated with an optional scannable barcode.

Barcode Workflow:

Use of barcodes for this SOP is highly recommended. Barcodes may improve sample tracking, and
reduce transcription errors associated with writing sample and subsample identifiers by hand. Barcodes
may also speed entry of data into the Herbaceous Clip Harvest Lab Masses app. Until they are linked
with a subsample, barcodes do not contain information specific to sample provenance.

When using barcodes, adhesive barcode labels should be applied to dry, room temperature bags or
envelopes in advance of their use in the field (at least 30 minutes prior, but may be applied at the start
of the season). Barcodes are unique, but are not initially associated with a particular sample; if using
barcodes, it is encouraged to apply these in advance. See Section Section A.3 of RD[04] for the
appropriate barcode label type for this procedure. Note that a barcode label is applied in addition to
labeling the subsample with human-readable information (hand-written or printed).

Barcodes are scanned into the mobile application when the subsample (i.e., herbGroup) is placed into
the bag; only one barcode may be associated with a particular subsample. Do not reuse barcodes. If a
barcode is associated with multiple subsamples, the data ingest system will throw an error and refuse to
pull in entered data.

SOP A

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SOP B Field Sampling Aboveground Biomass of Agricultural Crops

At agricultural sites, clip harvesting produces estimates of total crop biomass production on a per crop
basis, as well as the biomass production of any non-crop herbaceous plants growing with the crop. Crop
biomass is separated from non-crop biomass, and non-crop biomass is further sorted to herbGroup as in
RD[04] (Figure 4).

In the procedure below, it is assumed that technicians have a working knowledge of SOP B in the TOS
Protocol and Procedure: Measurement of Herbaceous Biomass (RD[04]).

Figure 4. Expanded workflow diagram for field sampling the aboveground biomass of agricultural crops (SOP B).
Diagram supports and does not replace protocol text; most common workflow is outlined.

SOP B

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HELPFUL HINT: COORDINATING WITH OTHER PLANT PROTOCOLS


Plant Diversity:
• If plant diversity sampling is scheduled to occur prior to crop clip-harvest sampling in a given
sampling year, it may be helpful to identify and demarcate a suitable Clip Strip prior to
performing diversity sampling. This will ensure that the Clip Strip is not trampled during diversity
sampling.
• Should clip-harvest occur before diversity sampling, take care to avoid trampling 1 m2 nested
subplots used for Plant Diversity % cover measurements.
• Reject the clip cell if rotation required for sampling causes the desired clip cell to overlap with
nestedSubplots used for Plant Diversity (applicable only in Distributed Plots and those Tower
Plots that support Plant Diversity).
Plant Belowground Biomass:
• Plant Belowground Biomass soil samples should be collected according to the standard protocol
from 0.5m x 0.5m sampling areas.
• Soil sampling areas for belowground biomass are rotated along with the crop clip strip and
‘book-end’ the clip strip on each narrow end, similar to standard North/South clip strips. See
Figure 3.

B.1 Sample Collection in the Field

1. Navigate to the selected plot, using the GPS if necessary. Should the farmer harvest the crop or
till a fallow plot prior to scheduled Agricultural Clip Harvest sampling:
• Create a record in the Site Management app describing the management activity that
occurred, and estimate the date of occurrence.

• If live, green weeds or other non-crop plants are still growing in the plot:
o Create a record for the clipID in the Herbaceous Clip Harvest Field Sampling app,
and indicate targetTaxaPresent = ‘Y’.
o In the remarks: Record ‘Crop harvest preceded NEON sampling’ or ‘Plot tilled
before NEON sampling’.
o Perform subsequent steps below.

• Else if no weeds or non-crop plants are growing:


o Create a record for the clipID, and indicate targetTaxaPresent = ‘N’.

SOP B

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o In the remarks: Record ‘Crop harvest preceded NEON sampling’ or ‘Plot tilled
before NEON sampling’.
o Proceed to the next plot.
2. As described in RD[04], use the plot- or subplot-specific Clip List to identify and locate the target
clip cell for sampling (steps 2 – 7 in RD[04]).
• When these steps are complete, you should have placed a pin flag at the SW corner of
the Clip Strip. This flag is referred to as ‘Flag1’ below (Figure 5). Because Ag Clip Strips
are commonly rotated, Flag 1 will not necessarily be the SW corner as it is for the
standard Clip Harvest.
• On the Clip List, record the clip cell that was used as the starting point for clip cell
delineation. Because agricultural clip strips are often oriented off the typical
north/south axis, they will typically intersect > 1 clip cell, and there is no need to
determine exactly which clip cells an angled clip strip intersects.
3. Select the appropriate crop-specific dimensions for the Clip Strip (Table 3).
4. Delineate the accepted Clip Strip for harvesting. Clip Strip orientation is not the same as in
RD[04]. For agricultural crops, clip strips are oriented perpendicular to crop rows in order to
consistently account for the spatial structure introduced by row planting (see Figure 5). Crop
row orientation, and therefore Clip Strip orientation, may change from year to year.
a. Stand over the top of ‘Flag1’ facing North, and rotate clockwise until you are
perpendicular to the crop rows. A compass may help ensure you are consistently
perpendicular to the rows.
b. Extend a meter tape or measuring stick outward to the required ‘Length’ (L) distance
specified in Table 3, and mark this distance with another pin flag – ‘Flag2’.
c. Stretch a string and stake set or lay the measuring stick between Flag1 and Flag2 to
mark one long edge of the Clip Strip.
Note: For crops in Table 3 that require Clip Strips with length < 2 m, use the folding
measuring stick instead of a string/stake set.
d. Standing over Flag1 again, look down the length of the string toward Flag2, and use the
meter tape or measuring stick to place two more pin flags X cm to the right of Flags1
and 2, where X is the ‘Width’ (W) distance (Table 3, Figure 5).

• A folding meter stick may be used to create an accurate right-angle help


determine the correct location of Flags 3 and 4.
e. Stretch a second string and stake set between these two new flags. The second string
should be parallel to the first, and the strings and flags should delineate a rectangular
Clip Strip with the dimensions shown in Table 3.

SOP B

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Figure 5. Clip Strip establishment in an agricultural row crop. Numbered flags correspond to those referenced
above in step (5); ‘Flag 1’ corresponds to the SW corner of the Clip Strip that is provided in the Clip List. Values for
the length (L) and width (W) of the Clip Strip are crop-specific and are provided in the Table 3 below.

Table 3. Clip Strip dimension required for crops expected at NEON agricultural sites. Row spacing is for typical
plantings and may vary from listed value.

Quadrat dimensions,
Crop Row spacing L x W (m)
Barley 18 cm (7 in) 2.0 x 0.5
Corn 38 or 76 cm (15 or 30 in) 1.5 x 0.65
*Millet 18 cm (7 in) 2.0 x 0.5
Sorghum 38 or 76 cm (15 or 30 in) 1.5 x 0.65
Soybeans 38 cm (15 in) 1.5 x 0.65
Wheat 18 cm (7 in) 2.0 x 0.5
* Millet is a generic term for a number of different crop plants. Submit a help ticket if
observed spacing is different from that listed here.

SOP B

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ROW SPACING

• If row spacing differs from that listed in Table 3 for a given crop by > 25%, submit a help ticket to
determine whether a different quadrat dimension is warranted.
• An early season reconnaissance may be helpful to detect potential crop spacing, and address
Clip Strip dimension issues prior to scheduled sampling.

5. Create a record in the Field Sampling app for the sampled plotID, and enter required plot-level
field sampling information:

• weekBoutBegan; the ISO week number in which sampling began. If a bout duration
exceeds a single week, enter the ISO number of the week the bout began. Multiple
bouts per season will be required if crop rotation is practiced at the site.
• collectDate; use YYYYMMDD format.

• clipCellNum; as provided in Clip List; use last 3 numbers from the clipID, e.g. for
BLAN_001_126, record ‘126’.
• clipDimension; The length (L) and width (W) of the Clip Strip, in meters; e.g., ‘1.5 x 0.65’.

• targetTaxaPresent; If there is no biomass in a Clip Strip (i.e., neither crop nor non-crop
biomass is present), AND the Clip Strip is deemed representative of the plot, record
‘targetTaxaPresent = N’
• hbpType; select ‘Agricultural’

• cropType; select the appropriate crop from the drop-down list. Submit an incident ticket
if the required crop type is not in the list.

• herbGroups; indicate ‘Present/Not present’ for all non-crop herbGroups. The Lab Mass
app will require a dryMass value for all ‘Present’ herbGroups.

SOP B

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6. Using a permanent marker, label an appropriately sized paper bag with the information below
(use large yard-debris paper bags for high-volume crops like corn). Remember that you will need
to clip and sort any non-crop biomass into the herbGroups listed in Table 10 of RD[04], so label
additional smaller 8# or 25# craft paper bags for this biomass, if present.
Note: To improve sample tracking, bags should be labeled with pre-affixed barcodes.
• weekBoutBegan; use ISO week format, e.g., ‘47’

• date; use YYYYMMDD format

• clipID; e.g. BLAN_001_126

• herbGroup; for crops, this will simply be the crop type (e.g., ‘corn’, ‘soy’, etc.), and for
non-crop biomass, herbGroup values are identical to those in RD[04].

• bagCount; the total # of bags generated from a given clipID (for Field Ops tracking
purposes only).
7. Clip and sort all crop and herbaceous non-crop biomass rooted within the Clip Strip. Recall that
an individual must be ≥ 50% rooted within the Clip Strip to be counted as ‘in’ for clipping.
General guidelines:
• For corn brace roots (Figure 1), clip all brace roots for an individual if it is ≥ 50% rooted
within the Clip Strip.
• Do NOT clip vegetation that passes through/leans over the Clip Strip but is not rooted in
the strip.
• DO include leaves and stems in the harvest that exit the strip, but originate from stems
rooted within the strip.
• When subsampling is required, use the Subsample Fresh Mass Target Calculator in the
(TOS) Herbaceous Clip Harvest: Field Sampling [PROD] application.

SOP B.1.1 Guidelines for high-volume crops with large seeds

• See Definitions (Section 2.4) for high- vs. low-volume crops, and note that it is always
preferable to avoid subsampling if possible. The high-volume procedure for crops with
large seeds is suitable for corn and other large-seeded crops.

• For steps that require the use of a spring-scale, always select the scale that allows the
greatest precision possible for the mass being weighed.
Step-by-Step Sub-sampling Procedure (Figure 6)

a. Prepare two bags:


i. A large bag for the entire fresh sample (yard waste type, no label required), and
ii. A smaller, labeled bag for the subsample (typically a #25 bag).

SOP B

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b. Tare bags on spring scales:


i. Tare both bags together; a 10 kg spring-scale is typically adequate for corn but
smaller scales may be necessary for low-yield plots.
ii. Tare the smaller subsample bag alone on a 5 kg spring-scale.
c. Separate ears or seed pods from the vegetative stalks/leaves, placing the ears/pods into
the smaller subsample bag and the stalks/leaves into the large bag.
i. If harvesting corn, leave ears intact.
ii. Use hand pruners or long-handled loppers to break up large stems and leaves.
d. Weigh the combined total mass of the content of both bags. To prevent tearing, double
over the top of the bag before passing the spring-scale hook through the paper. Bag
tops may also be re-inforced with tape and then hole-punched.
i. Record the Fresh Mass to the greatest precision afforded by the spring-scale
(nearest 50 g for a 10 kg scale).
e. Weigh the ears/pods separately on the 5 kg spring-scale.
i. Record the Seed Fresh Mass to the greatest precision possible.
f. Calculate the ‘Seed Mass:Fresh Mass’ ratio by dividing Seed Fresh Mass by Fresh Mass
(done automatically by the Calculator in the Fulcrum app).
g. Select a representative ear or handful of pods and leave in the small bag to weigh. The
remaining ears/pods may be discarded.
i. Record the Sub Sample Seed Fresh Mass to the greatest precision possible.
h. Calculate the target Subsample Fresh Mass by dividing the Sub Sample Seed Fresh
Mass by the Seed to Fresh Mass Ratio (done automatically by the Calculator in the
Fulcrum app).
i. Add representative handfuls of vegetative biomass from the large bag to the smaller
subsample bag until the target Sub Sample Fresh Mass is achieved (± 100 g).
i. Make sure the proportion of stalks/leaves is roughly maintained when adding
vegetative mass to the subsample bag.
j. Weigh and record the actual Sub Sample Fresh Mass to the greatest precision possible.
k. Place the subsample into cold storage as soon as possible (e.g., cooler with re-usable
cold packs), and transport back to the laboratory for drying.

SOP B

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Figure 6. Subsampling routine for high-volume crops with large seeds (e.g., corn). Letters (a-j) correspond to
protocol steps.

Example:

• The Fresh Mass of a corn Clip Strip is 5000 g (leaves/stalks + ears), and the Seed Fresh Mass is
2000 g (ears only). The Seed:Fresh ratio is 2000/5000 = 0.4

• One representative ear is selected for the subsample, with Sub Sample Seed Fresh Mass = 500 g.
The target Sub Sample Fresh Mass = Sub Sample Seed Fresh Mass / Seed:Fresh = 500/0.4 = 1250 g.
• Add vegetative mass to the existing ear until the total mass is 1250 ± 100 g. This is the Sub Sample
Fresh Mass to place in the cooler and take back to the laboratory.

SOP B.1.2 Guidelines for high-volume crops with small seeds

• See Definitions (Section 2.4) for high- vs. low-volume crops. The high-volume procedure for
crops with small seeds is suitable for grains and other small-seeded crops.

Step-by-Step Sub-sampling Procedure (Figure 7)

a. Prepare and label two bags as above for high-volume crops with large seeds.

SOP B

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b. Tare bags separately on spring-scales. Spring-scale sizes below are for typical plots; ; a
smaller scale may be necessary for low-yield plots:

i. Tare the large bag on a 10 kg spring-scale.


ii. Tare the smaller bag on a 5 kg spring-scale.

c. Place all clipped biomass into the large bag, and do NOT separate seed heads from
vegetative stalks/leaves.
d. Weigh and record the Fresh Mass to the greatest precision afforded by the spring-scale
(nearest 50 g for a 10 kg scale).
e. Create a subsample in the smaller bag and visually maintain the existing proportion of
seeds/leaves/stalks.
i. This is most easily done by selecting whole, representative plants for
subsampling.
ii. The subsample should be 20% – 30% of the initial Fresh Mass.
f. Weigh and record the Sub Sample Fresh Mass to the greatest precision possible (i.e., to
the nearest 25 g with the 5 kg spring-scale).

SOP B

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Figure 7. Subsampling routine for high-volume crops with small seeds (e.g., cereals). Letters (a-f) correspond to
protocol steps.

(1) Guidelines for low-volume crops (some cereals, etc.): Do not create a sub-sample
a. Place the entirety of the clip-harvested crop sample into the bag from step (7).
b. Place the bagged sample into cold storage as soon as possible, and transport back to the
laboratory for drying.
c. Select ‘Fresh Sub Samples Created’ = No
d. The Fresh Mass and Sub Sample Fresh Mass fields are not required (these fields are
automatically hidden by the ingest application).

8. Record the total number of bags from the Clip Strip on the bags, and record the time bags will
be placed in the cooler in the Field Sampling app (estimate to the nearest 30 min).
• A value for time is still required for the record even if there are no samples placed in the
cooler.
9. If using barcodes: Link barcodes from each bag with subsamples in the Field Sampling app
(subsamples = crop and herbGroups present).
10. Save the Field Sampling record. Records are finalized in a later step after sort-checking of non-
crop biomass is completed.
11. When clipping is finished, group any small 8# bags from non-crop samples from the clipID
together into a 25# bag, label with the clipID and date, and place in cold storage.
• If cold storage space is insufficient for the crop volume, seal paper bags containing the
sample, and keep at ambient temperature while still in the field.
12. If a high-volume crop was subsampled, dispose of any excess fresh biomass that will not be
brought back to the laboratory outside the plot.
13. Return to step (1) for the next plotID.

B.2 Sampling Non-Standard Agricultural Plots

Table 2. Additional guidelines for field conditions that required special handling or consideration.

Field condition Guidelines


Plot planted in more than Accept a random Clip Strip that falls within the dominant crop, as visually
one crop type assessed by % cover (Figure 8). Reject other Clip Strips. Note that rejected
Clip Strips are not rejected permanently as are un-representative Clip
Strips in RD[04], due to the fact that crop cover changes regularly (just
skip over these strips in the list).

SOP B

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Field condition Guidelines


Non-target crop is present Select cropType for the dominant, target crop (e.g., wheat). Assign the
but rare amongst target non-target crop to the appropriate herbGroup and treat as non-crop
crop – e.g., a few corn biomass.
stalks grow in a field of
wheat
Wind thrown corn with Only clip biomass associated with rooting points located within the strip.
multiple root points, at
least one of which is in the
Clip Strip
Stubble from previously If the plot was harvested by the farmer before the scheduled NEON Clip
harvested crop is present Harvest bout: Clip stubble only if stubble biomass was produced in the
(e.g., wheat stubble) current season, and note in remarks, “Clip occurred after farmer
harvest.”

Do NOT clip stubble produced in a previous growing season.


Rows cannot be discerned Visually assess the plot with respect to % cover (rows can vs. cannot be
for a portion of the plot discerned). If rows cannot be discerned for > 50% of the plot, then move
due to non-crop growth forward with a standard N/S clip strip, and reject cells where rows can be
discerned.

Soybean

Corn

Figure 8. Rough percent cover map of plot planted in both corn (dominant crop) and soybeans (subdominant crop).
The map may be used to reject potential clip strip locations that fall within the ‘soybean’ area without navigating
to them.

SOP B

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B.3 Sample Preservation

• Keep paper bags with clipped vegetation in a cooler with cold packs to minimize wilting and
biomass loss.

• Change cold packs for fresh ones every 12 h or transfer to a 4 ˚C refrigerator if a drying oven is
not immediately available.

• Return to the laboratory for drying within 24 h of clipping, if possible.

• Transfer bags of clipped biomass to the drying ovens as soon as possible after field sampling,
and monitor drying progress with the “Lab Drying” data sheet.

• Remember that samples must be dried within 5 days of clipping.

!!! IMPORTANT: Record the clipDate and time in the Herbaceous Clip Harvest: Field Sampling app AND
ovenInDate and time on both the sample bags and in the Herbaceous Clip Harvest: Lab Masses app so
that the number of hours the bags were stored cold can be automatically calculated.

SOP B

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SOP C Post-Field Sampling Tasks

The workflow following field work is largely similar to the analogous activity in the protocol (RD(04):
checking biomass bags and sorting (Figure 9).

Figure 9. Expanded workflow diagram for sort-checking field-collected agricultural biomass prior to oven-drying in
the laboratory (SOP C). Diagram supports and does not replace protocol text; most common workflow is outlined.

C.1 Sort-Checking non-Crop Biomass

1. Sort-check non-crop biomass to ensure accurate herbGroup classification. Use the procedure
outlined in RD[04].
2. Update and save herbGroup presence/absence in the Field Sampling [PROD] app (if necessary).

SOP C

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C.2 Refreshing the Sampling Kit

• Make sure the following consumables are available in sufficient quantity for the next round of
clip-harvests:
o Large ‘yard waste’ style paper bags, 30 gallon capacity (high volume crops). Bags may be
re-inforced with tape and hole punched at this time, if desired.
o Paper bags, 8# and 25# kraft (or the necessary size given site vegetation stature)
o All-weather paper for printing field datasheets
o Permanent markers for labeling bags in the field

• Return cold packs to the -20 ˚C freezer to refresh.

C.3 Equipment Maintenance and Cleaning

• Clean blades of hand clippers with an appropriate solvent (oil, ethanol, water), and dry
thoroughly.
• Recharge batteries for the GPS unit (if necessary).
• Recharge batteries for the TruPulse (if applicable).

C.4 Data Management

1. After sort-checking non-crop biomass (above), finalize and save all records in the HBP: Field
Sampling [PROD] app.
• Finalizing records will auto-generate corresponding records for each Clip Strip (parent)
and Herb Group (child) in the HBP: Lab Masses [PROD] app.
2. Sync all tablets. Tablets should be synced before any additional Lab Mass edits are made.
3. See RD[08] for additional Data Management guidelines that pertain to this procedure.

SOP C

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SOP D Laboratory Processing of Agricultural Biomass Samples

Overview

Oven dried ‘dryMass’ values are generated for crops and herbaceous non-crop plants from the same clip
strip. Drying and weighing of clip-harvested crop biomass is very similar to that described for
herbaceous biomass in RD[04], with the exception that high-volume crop biomass is subsampled prior to
oven drying. A subsampling approach ensures the drying ovens are not monopolized by a small number
of high-volume samples. Because of the subsampling approach used for high-volume crops, data are
entered into the ‘Lab Weighing’ ingest table differently for high-volume crop versus low-volume crop
and non-crop herbaceous biomass (Figure 10):

• High-Volume Crop biomass (SOP D.1):


o Values for Fresh Mass, and Sub Sample Fresh Mass are recorded in the field (SOP B).
o Record Sub Sample Dry Mass in the ‘Lab Weighing’ ingest application.
o Values of Dry Mass are calculated automatically.

• Low-Volume Crop and Non-crop Herbaceous Biomass (SOP D.2):


o Fresh Mass, Sub Sample Fresh Mass , and Sub Sample Dry Mass are left blank.
o Record measured Dry Mass of entire clipped sample.

SOP D

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Figure 10. Expanded workflow diagram for drying and weighing clipped, sort-checked agricultural biomass (SOP D).
Diagram supports and does not replace protocol text; most common workflow is outlined.

D.1 Drying and Weighing Clipped High-Volume Crop Biomass

Most of the crops to which this SOP applies will be high-volume crops, and clipped biomass from all
plots will therefore not readily fit into NEON’s drying ovens. Certain cereals may be low-volume crops; if
all of the clipped crop samples can fit into the drying ovens, use the low-volume crop biomass approach
(SOP D.2, below).

1. Place labeled 25# subsample bags into a 65 ˚C drying oven for 48h – 120h (2d – 5d).

• Some crop samples may take significantly longer to dry, make sure there is adequate
oven space.
2. After placing all bags from one clipDate in the oven, check the drying progress of clipped
biomass using a subset of 10 bags, and the “Lab Drying” datasheet.
a. Check the weight of the same selected subset of 10 bags per clipDate after day 1, 2, 3,
etc. Record these weights each day on the “Lab Drying” datasheet.

SOP D

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b. Calculate the difference in weight between the latest two time points for each bag.

c. Subsamples are dry when the average weight difference between the last two time
points = 0 (averaged across all 10 bags, ± 0.1 g or ± 1.0% of the previous timepoint mass,
whichever is larger).

AGRICULTURAL BIOMASS DRYING TIPS

• A spreadsheet calculator is useful for calculating the average weight difference. A link is
provided in the ‘Supporting Documents’ section of the Field Ops Sampling Support Library on
Sharepoint.
• To save time, plant material should be weighed WITH the bag to prevent loss of material during
drying.
• Focus on the heaviest bags, as these will likely take the longest to dry.
• Additional drying tips in RD[04] also apply.

3. Remove bags of dried biomass from the drying oven, and label bags with ovenOutDate/Time.
• Dried plant material should be weighed immediately after removing from the drying
oven, as it will absorb moisture from the air if left in ambient room conditions
(particularly in humid environments).
o If using this method, it is helpful to remove bags from the oven and weigh one
at a time.
• Dried subsamples may also be stored for up to 30 days in ambient room conditions prior
to weighing. Subsamples treated in this manner must be returned to the drying oven for
24 h prior to weighing, and must be weighed as above after removal from the oven.
o If samples have been initially dried and kept in storage, it is not necessary to
record any additional drying times.
4. Weigh crop subsamples from each clipID using an electronic scale, and a plastic tray or weigh
boat.
• If using optional barcodes:
o Open the Herbaceous Clip Harvest: Lab Mass app, and scan the barcode on the
bag to bring up the record associated with the plotID.
• Select the appropriate crop/herbGroup subsample that matches the bag, and choose
‘Edit’.
• Record Sub Sample Dry Mass; nearest 0.01 g, plant material ONLY (without the bag).
• Avoid splitting the subsample into subgroups for weighing, as uncertainty values from
weighing must be added each time a subgroup is created.
• Do NOT record Dry Mass. This will be calculated automatically by the Fulcrum ingest
application.

SOP D

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5. Record and/or check required metadata for each subsample in the “Lab Weighing” ingest table:
• weighDate; date subsample was weighed in the laboratory, YYYYMMDD format
• plotID; SITE_XXX format
• clipCellNum; last three digits of the clipID
• clipDate; date sample was clipped in the field, YYYYMMDD format
• ovenInDate / Time; date and time (24-h format) subsample was placed in oven (initial
drying only)
• ovenOutDate / Time; date and time subsample was removed from oven (initial drying
only)
• herbGroup; select ‘corn’, ‘wheat’, ‘soy’, ‘sorghum’, etc.

• Save the herbGroup level child record, and when all child-records have been updated
with Dry Mass values, save the parent plotID record.

D.2 Drying and Weighing Clipped Low-Volume Crop and Non-crop Herbaceous Biomass

Low-volume crop biomass and non-crop herbaceous biomass is dried and weighed as described in
RD[04]. In contrast to high-volume crop biomass processing above (SOP D.1), no subsampling is
employed here: Dry and weigh the entire sample, as in RD[04].

1. Enter required data into the Herbaceous Clip Harvest: Lab Mass app.

• If using optional barcodes:


o Open the Lab Masses app, and scan the barcode on the bag to bring up the
record associated with the plotID.
• Select the appropriate crop/herbGroup child record that matches the bag, and choose
‘Edit’.
• Record herbGroup; for low-volume crop, select ‘wheat’, ‘barley’, etc.; for non-crop
biomass select the appropriate herbGroup described in RD[04] (i.e., cool-season
graminoid, leguminous forb, etc.)
• Record Dry Mass; total dry mass of the entire sample, nearest 0.01 g, plant material
ONLY (without the bag).

• Save the herbGroup level child record, and when all child-records have been updated
with Dry Mass values, save the parent plotID record.

D.3 Data QA

• QA sample weighing is not performed for high-volume crop samples.


• For low-volume crop samples and non-crop herbaceous biomass, QA sample weighing is
performed as in RD[04].
• Record QA weight data to the nearest 0.01 g in the qaDryMass field of the “Lab Weighing”
ingest table.

SOP D

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SOP E Data Entry and Verification

The Aboveground Productivity for Agricultural Crops SOP uses the Herbaceous Biomass data entry and
verification workflow and ingest, and is identical to that described in SOP F of RD[04]. Consult the Data
Management protocol for Herbaceous Biomass data quality procedures (RD[08]).

SOP E

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APPENDIX A EQUIPMENT

The following equipment is needed to implement the procedures in this document. Equipment lists are
organized by task. They do not include standard field and laboratory supplies such as charging stations,
first aid kits, drying ovens, ultra-low refrigerators, etc.

Table 4. Equipment list – Equipment needed to prepare for sampling.

Supplier/ Exact
Description Purpose Quantity
Item No. Brand

DURABLE ITEMS

GPS receiver, decimeter accuracy


Navigate to sampling locations at
Compass (e.g. Trimble GEO XH 6000,
Y sites where plot markers are 1
Tools Trimble GEO 7X, or equivalent);
absent
Range pole, and antenna

Amazon
GPS receiver, recreational Navigate to sampling location at
Cabela’s N 1
accuracy sites with plot markers
REI

Forestry
TruPulse 360R Laser Rangefinder, Delineate plot boundaries,
Supplier Y 1
± 30 cm accuracy determine clip cell location

USB Cable Transfer data to GPS unit. 1

CONSUMABLE ITEMS

All weather paper Printing field datasheets 1

Labeling sample containers with


Adhesive barcode labels (Type I) 1 sheet
barcode-readable labels

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Table 5. Equipment needed for a 2-person team to perform agricultural clip harvest sampling at one plot.

Supplier/ Exact
Description Purpose Quantity
Item No. Brand

Durable Items

GPS receiver, decimeter accuracy


Compass (e.g. Trimble GEO XH 6000, Navigate to sampling locations at
Y 1
Tools Trimble GEO 7X, or equivalent); sites where plot markers are absent
Range pole, and antenna

Amazon
GPS receiver, recreational Navigate to sampling location with
Cabela’s N 1
accuracy plot markers
REI

Ben Locate clip-harvest strips within


Meadows plots/subplots; measure and
N Measuring tape, minimum 30 m 1
Forestry delineate dimensions of clip harvest
Supplier strip.

Ben
Meadows Compass with mirror and Locate clip-harvest strips (with
N 1
Forestry declination adjustment measuring tape)
Supplier

Forestry TruPulse 360R Laser Rangefinder, Delineate plot boundaries,


Y 1
Supplier ± 30 cm accuracy determine clip cell location

N Pruning shear Clip plants 2

N Loppers, long-handled Clip and cut corn 1

Forestry Spring scale, 10 kg capacity, Weigh total fresh mass of high-


N 1
Suppliers tareable volume crops (e.g., corn)

Forestry Spring scale, 5 kg capacity, Weigh subsample fresh mass of


N 1
Suppliers tareable high-volume crops (e.g., corn)

Spring scale, 2.5 kg capacity,


Forestry tareable. Note: Unit has English Weigh subsample fresh mass for
N 1
Suppliers and metric gradations. Data graminoid crops (e.g., wheat)
should be recorded in metric.

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Supplier/ Exact
Description Purpose Quantity
Item No. Brand

Durable Items

Spring scale 1000 g capacity,


Forestry tareable. Note: Unit has English
N Weigh low mass subsamples
Suppliers and metric gradations. Data
should be recorded in metric.

N Cooler Chill perishable samples in field 1

Fisher
N Cold packs Chill perishable samples in field
Grainger

Delineate clip harvest strip for small-


Pre-marked string and stake sets;
N stature crop types (measuring stick 2
see RD[04] for more details
may be easier if vegetation is thick)

Delineate clip harvest strip for crops


Forestry Measuring stick, minimum 2 m
N (may be easier to push through thick 1
Suppliers length, folding; or equivalent
vegetation than using string sets)

Ben
Meadows Chaining pins or other suitable
N Anchor measuring tapes 2
Forestry anchor
Suppliers

Survey marking flag, PVC or


N Delineate sampling area 4
fiberglass stake

N Work gloves Protect hands 2

Forestry Aid in species identification


N Magnifier hand-lens, 10X 1
Suppliers (particularly for non-crops)

Forestry Aid in species identification


N Magnifier hand-lens, 20X 1
Suppliers (particularly for non-crops)

Contain clipped herbaceous


ULINE N Paper bags, #81 62
biomass, sorted to functional group

Large paper bags, yard waste type


N or equivalent (e.g., 30 gallon Contain clipped crop biomass 1-42
capacity)

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Supplier/ Exact
Description Purpose Quantity
Item No. Brand

Durable Items

N Permanent marker Label paper bags 2

N CR123A battery Spare battery for laser rangefinder 2

N AA battery Spare battery for GPS receiver 2

Calculation of seed:fresh ratio when


N Field notebook 1
using paper data sheets.

Resources

Herbaceous Biomass Field


RD[05] NA Record sampling metadata
Datasheets

As
NA Per plot or subplot Clip Lists Identify random Clip Strip locations
needed

Field guide, regional flora Identify leguminous forbs and


NA 1
reference guide and/or key graminoids to species

Table 6. Equipment needed for post-field sampling tasks.

Supplier/ Exact Quan-


Description Purpose
Item No. Brand tity

Durable Items

Forestry Aid in non-crop species


Y Magnifier hand-lens, 10X 1
Suppliers identification

Forestry Aid in non-crop species


N Magnifier hand-lens, 20X 1
Suppliers identification

Cooler, size dependent on Chill perishable non-crop and low- As


N
anticipated crop volume volume crop samples needed

Chill perishable non-crop and low-


N Cold packs Variable
volume crop samples

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Supplier/ Exact Quan-


Description Purpose
Item No. Brand tity

Durable Items

Resources

Completed Herbaceous Biomass Contains field-collected sampling


RD[05] NA Variable
Field Datasheets metadata

Field guide, regional flora Aid in distinguishing morphologically


NA 1
reference guide and/or key similar species to functional groups

Table 7. Equipment needed for processing agricultural biomass clip harvest samples in the laboratory.

Supplier/ Exact Quan-


Description Purpose
Item No. Brand tity

Durable Items

Fisher N Balance, 0.01 g accuracy Weigh dried subsamples 1

Contain dried non-crop samples


Fisher N Weigh boats, large Variable
while weighing

Contain crop samples and


Fisher N Plastic tray or equivalent 1
subsamples while weighing

Consumable Items

Datasheets: As
Recording dry weight of herbaceous
RD[05] NA • Lab Drying QC Datasheet
biomass needed
• Lab Weighing Datashee

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NEON Doc. #: NEON.DOC.001714 Author: C. Meier Revision: E

APPENDIX B SITE-SPECIFIC CROP SAMPLING

B.1 D04 LAJA

Crops present (Distributed Plots):


• Mangoes (not sampled)
• Peppers (not sampled)
• Squash (not sampled)
• Sunflowers (sampled)
o Clip Strip dimensions: 1.5m x 0.65 m
o Crop Type: High-volume; see SOP B.1, step 8, section (1)
o Row Spacing: TBD
• Tomatoes (not sampled)
• Others (consult with Domain Manager and Science)

Distributed Plot sampling strategy:


• Only clip the those plots planted in pasture hay once per year in October, and keep the annual
clip harvest bout.
• Many of the plots planted with the above crops are located in small-scale experimental parcels
of an Agricultural Experimental Station, and support Master’s experiments, etc.
• Do not sample crops listed as ‘not sampled.’ Instead:
o Record `targetTaxaPresent = Y` for the plot/clipID.
o In the remarks, record ‘Experimental Crop: X, not sampled’, where X is one of the crops
listed immediately above.

Crops present (Tower Plots):


None: Tower Plots are grazed

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