Attachment 9- EXHIBIT H NEEDLE COLLECTION PROTOCOL Idaho.pdf

EXHIBIT H - Needle Collection Protocol for Identification of Whitebark and Limber Pine

Overview:

Whitebark and limber pine are ecologically important species that are impossible to distinguish in the field when cones

(male or female) are not present. We have access to an inexpensive genetic test that can distinguish between these two species (and only these two species) using needle samples. We will be collecting needles from whitebark and limber pine in areas where they are known to co-occur. Because this test only works to determine whitebark pine from limber pine, it is important that crews do not mistakenly send in other similar species (e.g., western white pine).

Where are we collecting needle samples?

These needle collection protocols should be followed by all crews working in counties listed at the end of this document

(also shown in the GREEN Area of interest map, below). This green area represents the known overlap of the area where limber and whitebark pines can potentially co-occur. If a plot is in one of the counties that intersects the green Area Of

Interest shown below, the PDR should prompt you for needle collection when the species for limber pine (113) or

1: The known ranges of whitebark (red), limber (blue) with a 35K buffer. The green area is the intersection of these buffered ranges. This green area is where mandatory needle collection for whitebark and limber pine seedlings, saplings, and trees without cones will occur.

whitebark pine (101) are entered for a seedling or sapling. All these counties fall within Montana, Nevada, Idaho, and

Wyoming.

It may be easier for you to note that needle sample collection is required on the front of your plot packets that are in these Area-of-Interest counties.

If you are outside of this area but suspect you might have a sample that may suggest a range expansion for whitebark pine or limber pine, please collect the sample and send it to the lab.

How do we know which trees (seedlings/saplings) to collect from on plots in these counties?

NEW AS OF 2023: You can use the lists linked below to get an idea of how many seedlings, saplings, and trees you should anticipate collecting needle samples for on the plot. Seedling and sapling estimates will be closer to what you end up collecting. Trees >5” DBH will be harder to accurately predict because collection is based on presence/absence of cones (see protocol below).

Seedlings: Collect samples for five seedlings of each species encountered on every microplot that is either whitebark or limber pine. For example, if you think you may have 8 whitebark seedlings and 10 limber seedlings on the subplot 4 microplot, collect samples from five of the seedlings you think are whitebark, then collect five more samples from those seedlings you think are limber pines. Try to collect samples evenly throughout the microplot (i.e. not from one cluster of seedlings).

Saplings: Collect needles from all live whitebark or limber saplings on the microplot.

Trees: Collect needles from all live mature (tally) whitebark or limber trees on plot when cones (male or female) are not present to determine the exact species. Read the attached PDF “Field Identification of Whitebark versus Limber Pine”

(also in the RMRS FIA FIELD Reference Library here) to find out more about how to tell these species apart.

Keep in mind that whitebark and limber can co-occur on the same plot, and they likely co-occur more frequently than we detect it. This means trees need to be identified on an individual basis; a definitive identification of one tree does not indicate a definitive ID for other trees on the subplot.

The genetic test can only differentiate between whitebark pine and limber pine.

Needle samples from other species may produce a false ID. Because of this, please be careful to send in only samples from whitebark or limber, when possible. Once again, if working in an area with other five-needle pine species, and you are unsure what 5-needle species a tree may be, you should still collect a sample, but make a note on the packet of what other species you think it may be.

What does a needle sample collection look like?

For healthy trees and saplings with relatively abundant needles: Collect a sample of six (6) intact needle fascicles (groups).

For seedlings and/or saplings with sparse foliage: Two full intact fascicle is preferred. If the seedling is extremely sparse, this sample size can be reduced to just half of a needle. Use professional judgement to not impact the health of the seedling.

Enter what you think the species is in the PDR (whitebark or limber pine). We will correct the data later, if necessary, when we get results back from the lab.

Guidelines for collecting samples:

https://usfs.box.com/s/s7la32e5p9czrhhtc7ngy8fqp9z2btov

• Use your best judgement and collect as many needles as you can (up to six intact fascicles) without damaging the individual.

• Try to keep the fascicles intact when collecting.

• If less than one fascicle is collected, please note this on the packet.

• Collect the greenest needles possible (e.g., with minimal non-green spots or fungal infections).

Put the needle sample in a small manila envelope, like the one shown on the right. Jim Sandall will mail these out to crews if you don’t already have them.

Pre-printed labels should be coming out soon, but in case you don’t have them, label the envelope as follows for each individual tree sample collected:

State – county – location – subplot - tree number (or "seed" if seedling) – species

If you collect multiple seedlings from a subplot, please identify how many were collected by labelling the envelopes ‘1 of

X’, ‘2 of X’, etc.

Within reason, do your best to store collection envelopes out of the heat in a dark, dry place while awaiting shipping to preserve the genetic integrity of the sample.

Please do NOT send these to your Area Leader or QA in the plot packet! Instead, once a month, mail them to Ogden. You can include your tree cores and plant samples as well.

Send all samples to:

Riverdale Forestry Sciences Laboratory

4919 S 1500 W, Ste. 200 Riverdale, UT 84405

Will there be follow up to prompt us to mail in samples or let us know whether we had correct species calls?

We plan to!

The plan is to send out notices one to two times throughout the season to crews that had whitebark and /or limber pine seedlings or saplings on plots in the Area of Interest, requesting that they mail in their samples.

We will also do our best to get information back to crews individually on where they were correct/incorrect on their identification of these seedlings/saplings.

Why are we doing this?

Identifying 5 needle pines in general, but particularly whitebark pines v. limber pines, is a known problem in botany/ecology. Botanists have determined that there is no way to definitively distinguish whitebark from limber pine when cones or other signifiers are not present (male or female). RMRS - FIA is no different.

Study results from a previous needle sample collection (2010) showed that in the Area of Interest, our species identification between whitebark and limber pine was accurate only about 80% of the time. Our accuracy was worse for seedlings and where both species co-occurred on the same plot.

Sending needles in to get genetically verified doesn’t cost much (about $2.50 per sample) and it increases our data quality for an important species (whitebark pine) that was listed as a threatened species by the US Fish and Wildlife

Service in December 2022.

FIA data are playing an important role in the legal status of this species. For example, FIA data were cited in the 2022 US

Fish and Wildlife Service’s decision, in the Federal Register, to list whitebark pine as a threatened species. (The “more than half of all standing whitebark pine trees are dead” soundbite, which is entirely based on FIA data, got a LOT of press!). On top of all that, limber pine is having its own struggles that warrant close tracking. Since FIA data is increasingly being used for making management decisions for these species, it only makes sense to provide the best data we can for these important efforts.

Counties with mandatory whitebark /limber seedling/sapling needle collection

State County State County State County State County

16 3 30 1 32 5 56 3

16 7 30 3 32 7 56 13

16 11 30 7 32 9 56 17

16 13 30 9 32 11 56 23

16 15 30 13 32 19 56 29

16 19 30 15 32 21 56 35

16 23 30 23 32 29 56 37

16 25 30 27 32 31 56 39

16 29 30 29 32 33

16 33 30 31 32 510

16 35 30 35

16 37 30 37

16 39 30 39

16 41 30 41

16 43 30 43

16 49 30 45

16 51 30 47

16 59 30 49

16 63 30 51

16 65 30 53

16 67 30 57

16 73 30 59

16 81 30 63

16 83 30 65

16 85 30 67

30 73

30 77

30 81

30 89

30 93

30 95

30 97

30 99

30 101

30 107

30 111

List of Plots with Possible Needle Sample Collection (Idaho)

State County

Code County Plot

Estimated Number of

Samples Idaho 39 Elmore 8 7

Idaho 49 Idaho 44 19

Idaho 49 Idaho 322 4

Idaho 49 Idaho 640 3

Field Season 2025

Identifying Whitebark versus Limber Pine in the Field

In RMRS-FIA, whitebark and limber pines are thought to co-occur in Montana, Nevada, central Idaho, and western Wyoming. Currently, whitebark pine is not known to grow in Utah, Colorado, Arizona, or New Mexico. See this guide, High Elevation White Pines, to view coarse-scale maps of approximate geographic distributions for both limber and whitebark pines. Click on these links for a finer-scale look at whitebark and limber pine predicted distributions. (Note: do not rely on range maps as definitive sources on the distribution of tree species.)

Cones: The easiest, most obvious, and accurate way to tell the difference between whitebark and limber pine is by their cones. If a tree is producing cones, they can be seen in the top canopy. Typically, old and occasionally chewed-off branches of fresh whitebark pine cones can be found scattered on the forest floor close to the trunk. Whitebark cones are deep purple in color, squat, thick, and sappy. They are extremely tight and difficult to open when they are fresh.

Whitebark cones are 2-3 inches in length whereas limber cones are 3- 8 inches long. Whitebark cones are purple and turn dark brown and don't flare open when mature. Limber cones are green and turn grayish brown and flare open when mature. Whitebark pine has a two-year cone.

This means, miniatures of the mature cone (about 1 inch long) appear in the first year, with the cone growing to maturity in the second year. Due to the fact that they are highly sought after by wildlife, intact, fresh whitebark cones rarely remain in the canopy once they have matured. Often, the remains or skeleton of the outer shell of a cone can be seen in the canopy after birds have eaten the fleshy seeds from the inside.

In contrast, limber pine cones are green in color and are significantly longer and thinner than whitebark. Because they are less sought after by wildlife, they tend to remain in the canopy, gradually turn brown, open their bracts, and eventually fall to the ground. It is more typical to find limber pine cones on the forest floor at the base of the tree than whitebark cones.

If cones are not visible, there is a period of time (early spring through late July-mid August) that the young male “catkin-like” cones or strobili, can be found on the terminal ends of the branches of both species. Young trees will begin producing these male pollen cones (strobili) many years before they produce any female pollen cones, making this a useful tool for smaller, younger whitebark pines.

The young whitebark male cones (strobili) and female cones are all bright red to deep purple, whereas the young limber pine cones (both male and female) are generally green-yellow (note: the exception to this is that for a very short time in early spring, the male

1. Whitebark pine cones. Photo courtesy of Lisa Landenburger from in the Interagency Whitebark Pine Monitoring Protocol for the Greater Yellowstone Ecosystem.

2. Limber pine cones. Photos courtesy of Lisa Landenburger from in the Interagency Whitebark Pine Monitoring Protocol for the Greater Yellowstone Ecosystem.

3. Early season "catkin-like" cones (aka, strobili) of whitebark pine. Note the bright pink-purple color. Photo courtesy of Karla Sartor from in the Interagency Whitebark Pine Monitoring Protocol for the Greater Yellowstone Ecosystem https://www.fs.fed.us/rm/highelevationwhitepines/About/dist.htm https://www.conifers.org/pi/Pinus_albicaulis.php https://www.conifers.org/pi/Pinus_flexilis.php https://irma.nps.gov/DataStore/DownloadFile/434245 https://irma.nps.gov/DataStore/DownloadFile/434245 https://irma.nps.gov/DataStore/DownloadFile/434245 https://irma.nps.gov/DataStore/DownloadFile/434245 https://irma.nps.gov/DataStore/DownloadFile/434245 https://irma.nps.gov/DataStore/DownloadFile/434245 https://irma.nps.gov/DataStore/DownloadFile/434245 https://irma.nps.gov/DataStore/DownloadFile/434245 https://irma.nps.gov/DataStore/DownloadFile/434245 immature cones of limber pine can also appear red). Later in the season (mid August- September), the whitebark “catkin- like” male strobili will begin to turn dark brown as they age. The limber pine “catkin-like” strobili begin to turn a more tan color as they age.

In a situation where a mixed stand is encountered, use cone identification as the main tool to identify between the two species.

Next, look at the immature female/male cones on the ends of the branches. Keep in mind the “note” mentioned above: If you encounter a tree that has just a few red “catkins” and green-yellow “catkins”, it is most likely limber pine. Make sure you survey the tree entirely before you draw any conclusions as to which species the tree belongs. If it is still unclear as to which species of conifers you are encountering and you cannot, with 100% confidence, collect a sample of six needle fascicles (groups) as described at the beginning of this document, put them in a properly labeled envelope, and send the sample to Ogden for genetic identification.

Elevation, substrate, and growth habit all provide clues to aid in the identification process, but none of these should be used solely to differentiate whitebark and limber pine. Whitebarks tend to fare better at the highest elevations, while limber pines are more common than whitebarks at lower elevations 6,000 feet and below. Both species can co-occur at higher elevations. Limber pines tend to do better than other tree species on limestone parent material, whereas whitebark is more common on granitic soils, especially where Luzula hitchcockii is present. Whitebark often grow in clumps because they frequently originate from Clark’s nutcracker seed caches. While less common, limber pines can also be cached by Clark’s nutcrackers and originate in clumps.

Presence or absence of cone-bearing trees nearby is not necessarily an indicator of presence/absence of whitebark pine or limber pine. Both species (but especially whitebark) are propagated by the Clark’s nutcracker, a bird that

5. This clumpy growth form, with multiple stems originating from one location, is common for whitebark pines. This occurs because their closed cones are harvested by Clark's nutcrackers.

The birds then disperse the seeds in small caches. The bird’s unretrieved caches become the new cohort of trees. While the Clark’s nutcracker does demonstrate a preference for whitebark pines, they also cache limber pines. This means that clumping alone is not a reliable indicator that a given tree is a whitebark.- R.Simons photo

4. Whitebark pine strobili that are beginning to fade, with white pine blister rust canker in the background. R. Simons.

will fly long distances to cache seeds-and by squirrels, which also carry seeds some distance from the source. Whitebark pine cones do not open and typically do not propagate by seeds falling out of cones, whereas limber pine cones do open and do propagate from nearby limber pines. Due to the existence of an aerial seed transport mechanism (birds), a nearby seed source is not required to encounter whitebark or limber pines.

References and additional resources:

Adapted from Interagency Whitebark Pine Monitoring Protocol for the Greater Yellowstone Ecosystem and resources listed here:

https://irma.nps.gov/DataStore/DownloadFile/434245 https://www.mdpi.com/1999-4907/10/12/1060/pdf https://www.fs.fed.us/rm/highelevationwhitepines/About/id.htm https://www.fs.fed.us/rm/highelevationwhitepines/About/dist.htm https://www.fs.fed.us/rm/pubs_int/int_gtr253.pdf https://rngr.net/publications/assisted-migration/whitebark-pine-operational-cone-collection-instructions-and-seed-transfer-guidelines https://oregonstate.edu/trees/conifer_genera/spp/pine_spp.html http://fwp.mt.gov/mtoutdoors/HTML/articles/portraits/whitebarkpine.htm http://dendro.cnre.vt.edu/dendrology/syllabus/factsheet.cfm?ID=227 http://dendro.cnre.vt.edu/dendrology/syllabus/factsheet.cfm?ID=176 https://www.conifers.org/pi/Pinus_flexilis.php https://www.conifers.org/pi/Pinus_albicaulis.php Thanks to Erin Shanahan from the Greater Yellowstone Inventory and Monitoring network in Bozeman, MT for help reviewing and revising this document.

https://irma.nps.gov/DataStore/DownloadFile/434245 https://irma.nps.gov/DataStore/DownloadFile/434245 https://www.mdpi.com/1999-4907/10/12/1060/pdf https://www.fs.fed.us/rm/highelevationwhitepines/About/id.htm https://www.fs.fed.us/rm/highelevationwhitepines/About/dist.htm https://www.fs.fed.us/rm/pubs_int/int_gtr253.pdf https://rngr.net/publications/assisted-migration/whitebark-pine-operational-cone-collection-instructions-and-seed-transfer-guidelines https://rngr.net/publications/assisted-migration/whitebark-pine-operational-cone-collection-instructions-and-seed-transfer-guidelines https://oregonstate.edu/trees/conifer_genera/spp/pine_spp.html http://fwp.mt.gov/mtoutdoors/HTML/articles/portraits/whitebarkpine.htm http://dendro.cnre.vt.edu/dendrology/syllabus/factsheet.cfm?ID=227 https://www.conifers.org/pi/Pinus_flexilis.php https://www.conifers.org/pi/Pinus_albicaulis.php