Forest Genetics Council Of

OTIP Application Form for 2017/2018

Name of Applicant/Project Leader Annette van Niejenhuis
Legal Name of Organization: Western Forest Products Inc.
Mailing Address: 8067 East Saanich Road, Saanichton, BC V8M 1K1
Telephone: 250.652.4023
Fax: 250.652.2800
E-Mail Address:
Financial Officer: Martin Hammond
Project number (provide the existing number if this is a recurring project, otherwise leave blank):
01 – WF2
Project Title:
Coastal Douglas-fir pollen contamination study
Outline of Project (use attached outline)
Douglas-fir crop genetic worth is estimated using a pollen contamination model that has not been fully verified using genomics tools. The current method results in reductions in estimated genetic worth of 25% compared to orchard contribution average in large crop years. The successful implementation of this project will result in a firm critique of our methods, improved estimates of genetic worth, and possibly a significant rise in estimated genetic worth of our larger crops.
Seed Planning Unit
01 / Annual SPU Seedling Need: / Years seed GW > 75% of potential / Orchard information (list all orchards considered in this application
Orch # / Avg. ramet age / GW / Avg. ramet
ht. (m) / # ramets in orch.
Species
Fdc / Seed Zone
M / Elevation band
1-900m / 15.3 / 0.2 / 197 / 18 / 2622
166 / 17 / 891
199 / 20 / 1810
Project Duration: 3 years / Project year: Year 3
Total Project Cost requested for FY 2017/18: $20,002.50
Anticipated future budget needs:
FY 2018/19: $ / FY 2019/20: $ / FY 2020/21: $


Project Description

Needs:

Douglas-fir, our leading coastal species for seedling demand, requires estimates of pollen contamination to determine the genetic worth of a crop. This is a registration requirement of the CF Standards for Seed Use, specific to coastal Douglas-fir, because production occurs where wild pollen of significantly lower genetic value is proven to be abundant. Pollen contamination estimates have significant effects on the registered genetic worth of a seed lot.

Estimates based on pollen monitoring range from 5% to 50% or more, depending on relative abundance of pollen counted on orchard and regional pollen monitors. At 50% pollen contamination, the genetic worth of a crop is reduced by 25%. This will amount to reducing the registered genetic worth of a crop from GW20 to GW15, if the weighted average breeding value of the parents contributing to the crop before pollen contamination is 20.

Estimates based on DNA sampling have been undertaken for a limited number of crops (reported by Webber, 2013).To date, three crops have been reviewed using genetic markers. In two of the three crops, genetic markers and pollen contamination monitoring indicated contamination rates that were similar and under 25%. In the third crop, a large crop, pollen monitoring indicated contamination above 45% but genetic markers suggested a much lower rate (under 20%). It is apparent that more crops need to be tested to determine the value of our pollen contamination estimates.

Pollen monitoring techniques are open to criticism: Orchard pollen is captured by monitors established within the bounds of the orchard. Pollen from outside sources is assumed to be captured in addition to orchard pollen. Regional monitors, placed a distance away from any of our Douglas-fir orchards, but on the Saanich Peninsula, are assumed to capture only non-orchard pollen. If orchard pollen shed begins a few days after regional shed has begun, reasonable estimates may be possible. As pollination occurs in early spring, cool damp weather is common, thus frequently the local population and orchard are in synchrony. The range of pollen movement from outside sources is not known. Do we get distant pollen included in our monitoring? If monitoring is picking up distant pollen sources, its viability may be negligible. Bohrerova et al (2009) have shown that the viable range of pine pollen is less than its dispersal range.

The Forest Genetics Council will be well-served to publish high genetic worth of seed crops, as this denotes success of the program and supports the Council’s volume gain objective. But these scores must be appropriate, and not an over-estimation of the growth potential of seed lots. At present, it appears that large crops may suffer from overestimation of pollen contamination, leading to underestimation of genetic worth. Significant effort and investment has been put into examining our protocol, but additional work is needed to improve our understanding and to support any protocol adjustments.

Zuzana Bohrerova, Gil Bohrer, Ki Don Cho, M. Adam Bolch, and Karl G. Linden 2009. Determining the viability response of pine pollen to atmospheric conditions during long-distance dispersal. Ecological Applications 19:656–667. http://dx.doi.org/10.1890/07-2088.1.

Objectives:

The objective of this project is to review our estimates of genetic worth by evaluating the pollen contamination factor. Three crop years with crops from multiple orchard sites will be assessed using genetic markers, and compared to standard contamination estimation practices. As a result of this work and work previously completed by Webber, and by El-Kassaby and team, we will verify our existing pollen contamination estimation practices or recommend changes.

Procedure:

The CFS lab and David Dunn will be contracted to determine the paternity of the seed provided, using proven genetic markers for Douglas-fir:

Up to 3 orchards will be included in the study. Eligible orchards will be located on the Saanich Peninsula, and will provide pollen monitoring data for the orchard(s) and crop-year(s) of interest. At least three years’ crops will be evaluated.

Year 1 (15/16)

Participating orchards will provide collections of 50 seeds from ≥5 healthy cones from 3-4 unique ramets, labelling each collection with a unique label (Orchard #, Clone #, map location).

Additionally, orchards will supply a bulked foliar sample for each pollen-producing clone present in the orchard (one ramet per clone for small trees; multiple copies for large ramets with significant pollen load), labelling each foliar sample (Orchard #, Clone #). It is expected that a very small number of labelling errors may become evident in the genetic fingerprinting of the bulked samples. Further exploration to determine source of error will be made for these clones.

Seed will be shipped to the CFS lab, where 25 filled seed per collection will be fingerprinted using genetic markers (250-300 seeds). Additionally, bulked foliar samples of each producing clone across all contributing orchards will be fingerprinted using the same genetic markers.

If not enough suitable crops are available, or if fewer orchards participate, we may select past crops that are not yet examined (2014, 2012) where pollen monitoring data is complete.

Data will be reviewed by the Douglas-fir breeder, and a progress report will be compiled.

Progress Two orchards provided seed from the 2015 crops for DNA analysis.

(See Summary at end of document.)

Year 2 (16/17)

We will repeat year 1 with additional crops. A progress report will be compiled.

Progress Three orchards provided seed from the 2016 crops and foliar samples for DNA analysis. Foliar samples are a one-time collection, and needed to be collected in year 2 because the appropriate timing for collection is shortly after bud break; we had not established the agreement with the lab early enough in year 1. Preliminary results of the DNA fingerprints are being reviewed.

(See Summary at end of document.)

Year 3 (17/18)

We will repeat year 1 with additional crops.

In year 3, a qualified genetics data analyst will be contracted to work with the Douglas-fir breeder to deliver genetic marker estimates of pollen contamination. The results will be combined and contrasted with work done by El-Kassaby lab at UBC and compared to existing estimation practices. Recommendations for improvement will be developed in consultation with orchard managers. A final report will be delivered.

Location:

Saanich Peninsula Orchards: Timberwest, MFLNRO, and WFP

Genomics Lab (CFS)

Output and Deliverables:

1.  Verification and recommendations for improved estimations of pollen contamination and subsequent genetic worth estimates of Douglas-fir seed crops produced on the Saanich Peninsula.

Budget:

In-kind support includes the provision and extraction of seed samples by orchards, and pollen monitoring data (funded, in part, by other OTIP projects).

Who will benefit from this work?

1. Coastal Licensees

2. Province of British Columbia

Project Team:

Carol Halldorsen Bevin Wigmore

Seed Orchard Technician TimberWest Orchard Manager,

Ministry of Forests, Lands, and Natural Resource Operations Mt. Newton Seed Orchard

Michael Stoehr Annette van Niejenhuis

Research Scientist, Douglas-fir Breeding Tree Improvement Forester

Tree Improvement Branch Western Forest Products Inc.

BC Ministry of Forests, Lands, & Natural Resource Operations Saanich Forestry Centre

Saanich Seed Orchard, 7380 Puckle Road 8067 East Saanich Road

SAANICHTON, BC V8M 1W4 SAANICHTON, BC V8M 1K1

250.652.7613 250.652.4023 ext. 2

Signature Block:

Name: Annette van Niejenhuis

Signature:

Date:

Progress Summary

Fdc Pollen Contamination Study / # / #extracted / Microsatellite Analysis completed (#/sample)
Foliar samples / 206 / 206 / 4
Germinated seeds from 2015 / 65 / 62 / 3-4
Germinated seeds from 2016 / 153 / 91 / 3-4
Seeds incubating/to be germinated (2016) / 176
Total samples for 2015 and 2016 / 600

All germinated Fdc seeds have been dissected into both tissue types, mega removed and stored frozen for possible future use, DNA extraction/analysis on remaining tissue. A subsample of mega tissues (36 samples) have been extracted and will be characterized to compare with foliar mothers.

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WFP01 DNA