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Evaluation of Cold—Moist Stratification Treatments for Germinating Eastern and Carolina Hemlock Seeds for Ex Situ Gene Conservation Author(S): Robert M

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Evaluation of Cold—Moist Stratification Treatments for Germinating Eastern and Carolina Hemlock Seeds for Ex Situ Gene Conservation Author(S): Robert M Evaluation of Cold—Moist Stratification Treatments for Germinating Eastern and Carolina Hemlock Seeds for ex Situ Gene Conservation Author(s): Robert M. Jetton, W. Andrew Whittier and William S. Dvorak Source: Southeastern Naturalist, 13(sp6):168-177. Published By: Eagle Hill Institute URL: http://www.bioone.org/doi/full/10.1656/058.013.s611 BioOne (www.bioone.org) is a nonprofit, online aggregation of core research in the biological, ecological, and environmental sciences. BioOne provides a sustainable online platform for over 170 journals and books published by nonprofit societies, associations, museums, institutions, and presses. Your use of this PDF, the BioOne Web site, and all posted and associated content indicates your acceptance of BioOne’s Terms of Use, available at www.bioone.org/page/ terms_of_use. Usage of BioOne content is strictly limited to personal, educational, and non-commercial use. Commercial inquiries or rights and permissions requests should be directed to the individual publisher as copyright holder. BioOne sees sustainable scholarly publishing as an inherently collaborative enterprise connecting authors, nonprofit publishers, academic institutions, research libraries, and research funders in the common goal of maximizing access to critical research. 2014 Forest Impacts and Ecosystem EffectsSoutheastern of the Hemlock Naturalist Woolly Adelgid in theVol. Eastern 13, Special US Issue 6 R.M. Jetton, W.A. Whittier, and W.S. Dvorak Southeastern Naturalist 6SHFLDO,VVXH ± (YDOXDWLRQRI&ROG±0RLVW6WUDWL¿FDWLRQ7UHDWPHQWVIRU *HUPLQDWLQJ(DVWHUQDQG&DUROLQD+HPORFN6HHGVIRU ([6LWX*HQH&RQVHUYDWLRQ Robert M. Jetton1,*, W. Andrew Whittier1, and William S. Dvorak1 $EVWUDFW Populations of Tsuga canadensis (Eastern Hemlock) and Tsuga caroliniana (Carolina Hemlock) are declining due to infestation by Adelges tsugae (Hemlock Woolly Adelgid), an exotic insect pest. A better understanding of the environmental conditions UHTXLUHGIRUVHHGJHUPLQDWLRQLVQHHGHGWRPRUHHI¿FLHQWO\XWLOL]HWKHVHHGVFROOHFWHGIRU genetic-resource conservation and the establishment of seed orchards. This study examined WKHHIIHFWRIFROG±PRLVWVWUDWL¿FDWLRQWUHDWPHQWVRIYDU\LQJGXUDWLRQ DQGGD\V RQWRWDOJHUPLQDWLRQ DQGWKHQXPEHURIGD\VWR¿UVWDQGSHDNJHUPLQD- tion (germination speed) on seeds of both species in experiments conducted at 22 °C and KKOLJKWGDUNSKRWRSHULRG2YHUDOOWRWDOJHUPLQDWLRQIRU(DVWHUQ+HPORFNZDV DQGLQFUHDVHGZLWKLQFUHDVLQJGXUDWLRQRIWKHVWUDWL¿FDWLRQWUHDWPHQWV&DUROLQD+HPORFN WRWDOJHUPLQDWLRQZDVDQGYDULHGOLWWOHDPRQJWKHWUHDWPHQWVDOWKRXJKIHZHUVHHGV WHQGHG WR JHUPLQDWH IROORZLQJ ORQJHU GXUDWLRQV RI VWUDWL¿FDWLRQ 6WUDWL¿FDWLRQ LQFUHDVHG germination speed of Eastern Hemlock but not Carolina Hemlock. The results indicate WKDW(DVWHUQ+HPORFNVHHGVVKRXOGEHFROG±PRLVWVWUDWL¿HGDW&IRUDWOHDVW±GD\V prior to sowing to promote higher total germination. Carolina Hemlock seeds can be sown GLUHFWO\IROORZLQJDKVRDNZLWKQRDGGLWLRQDOFROG±PRLVWVWUDWL¿FDWLRQ ,QWURGXFWLRQ Tsuga canadensis (L.) Carr. (Eastern Hemlock) and T. caroliniana Englem. (Carolina Hemlock) are long-lived, slow-growing, shade-tolerant conifers native to eastern North America. Eastern Hemlock occurs across a broad elevation range IURPVHDOHYHOWRPDQGKDVDZLGHVSUHDGJHRJUDSKLFGLVWULEXWLRQWKDWH[WHQGV from Nova Scotia west to northern Minnesota and south through New England, the Middle Atlantic States, and the southern Appalachian Mountains into northern *HRUJLDDQGWKH&XPEHUODQG3ODWHDXRI$ODEDPD )DUMRQ 7KHVSHFLHVLVEL- modal in habitat distribution, occurring in high abundance on moist, well-drained, QXWULHQWULFK VRLOV RI PHVLF ULSDULDQ ]RQHV DQG VHDVRQDOO\ PRLVW VXE[HULF DUHDV DWWKHORZDQGPLGGOHSRUWLRQVRILWVHOHYDWLRQUDQJH .HVVHOO $WKLJKHU elevations, it is often more scattered along exposed xerophytic slopes and ridges. Carolina Hemlock has a much smaller distribution and is restricted to the south- eastern United States where it is endemic to the southern Appalachian Mountain and Upper Piedmont regions of Virginia, Tennessee, Georgia, North Carolina, and 6RXWK&DUROLQD -HWWRQHWDO 8QOLNH(DVWHUQ+HPORFN&DUROLQD+HPORFNLV 1Camcore, Department of Forestry and Environmental Resources, North Carolina State 8QLYHUVLW\&DPSXV%R[5DOHLJK1& *Corresponding author - rmjetton@ ncsu.edu. Manuscript Editor: Scott Markwith 168 2014Southeastern Naturalist Vol. 13, Special Issue 6 R.M. Jetton, W.A. Whittier, and W.S. Dvorak PRVWDEXQGDQWDORQJGU\QRUWKIDFLQJURFN\ULGJHWRSVDWHOHYDWLRQVRI± PRQVRLOVWKDWDUHGU\DFLGLFDQGQXWULHQWSRRU +XPSKUH\ Both Eastern and Carolina Hemlocks are considered to be at-risk species, and a number of factors threaten their long-term sustainability in eastern North America %HDUGPRUHHWDO)DUMRQHWDO 7KHPRVWVHULRXVRIWKHVHWKUHDWVLV the invasive Adelges tsugae Annand (Hemlock Woolly Adelgid [HWA]), an exotic, aphid-like insect introduced from Japan into the eastern United States in the early VWKDWFXUUHQWO\LQIHVWVDSSUR[LPDWHO\RIWKH(DVWHUQ+HPORFNUDQJHDQG RIWKH&DUROLQD+HPORFNUDQJH 86'$)RUHVW6HUYLFH +:$IHHGV at the base of hemlock needles by inserting its piercing/sucking mouthparts and extracting stored nutrients from xylem ray-parenchyma, thereby disrupting vegeta- tive and reproductive bud formation, causing needle desiccation and defoliation, DQGNLOOLQJWUHHVLQ±\HDUV <RXQJHWDO $OWKRXJKHVWLPDWHVRIWKHQXP- EHURIKHPORFNVNLOOHGE\+:$LQHDVWHUQ1RUWK$PHULFDQDUHGLI¿FXOWWRGHULYH widespread decline and morality have occurred among populations of both native hemlock species and continues unabated in most areas. The integrated strategy to manage the impacts of HWA on hemlock ecosystems in eastern North America includes a cooperative genetic-resource conservation SURJUDPEHLQJFRQGXFWHGE\&DPFRUH ,QWHUQDWLRQDO7UHH%UHHGLQJDQG&RQVHUYD- tion Program) at NC State University, Raleigh, NC, and the USDA Forest Service Forest Health Protection, Asheville, NC. The objectives of this effort are to improve the general understanding of native hemlock genetic diversity, climatic and edaphic adaptability, reproduction and regeneration ecology, and silvicultural options for VHHGRUFKDUGHVWDEOLVKPHQWDQGWRXWLOL]HWKLVLQIRUPDWLRQWRGHVLJQDQGLPSOHPHQW ex situ germplasm conservation strategies for both Eastern Hemlock and Carolina +HPORFN -HWWRQHWDO 7KHSUHVHQWVWXG\DGGUHVVHVRQHDVSHFWRIKHPORFN UHSURGXFWLYHELRORJ\WKHHIIHFWRIFROG±PRLVWVWUDWL¿FDWLRQWUHDWPHQWVIRUDOOHYLDW- ing seed dormancy and improving germination. Hemlocks have male and female strobili that develop on the same branches (monoecious), and these are well distributed throughout the crown in open-can- opy conditions and are more restricted to the upper crown under closed canopies %DUERXUHWDO +HDY\FRQHFURSVLQQDWXUDOVWDQGVRFFXUDW±\HDULQWHU- YDOV *RGPDQDQG/DQFDVWHU0HDQV3DFNHH 6HYHUDOVWXGLHV have evaluated cold stratification treatments for improving germination of hem- lock seeds collected from natural stands for a number of Tsuga species, including (DVWHUQ+HPORFN %DOGZLQ6WHDUQVDQG2OVRQ T. heterophylla 5DI 6DUJ :HVWHUQ+HPORFN $OOHQ(GZDUGV(GZDUGVDQG2O- VHQ /L DQG %XUWRQ DQG T. mertensiana (Bong.) Carr. (Mountain +HPORFN (GZDUGVDQG(O.DVVDE\(O.DVVDE\DQG(GZDUGV 6WUDW- ification treatments to improve seed germination in Carolina Hemlock have not been studied previously. 7KHREMHFWLYHRIWKLVVWXG\ZDVWRGHWHUPLQHWKHHIIHFWVRIFROG±PRLVWVWUDWL¿FD- WLRQWUHDWPHQWVRIYDU\LQJGXUDWLRQ ±G RQWRWDOJHUPLQDWLRQDQGWKHQXPEHU RIGD\VWRUHDFK¿UVWJHUPLQDWLRQDQGSHDNJHUPLQDWLRQIRUQDWXUDOVWDQGVHHG VRXUFHVRI(DVWHUQ+HPORFNDQG&DUROLQD+HPORFN VHHGVRXUFHVSHUVSHFLHV 169 2014 Southeastern Naturalist Vol. 13, Special Issue 6 R.M. Jetton, W.A. Whittier, and W.S. Dvorak The data will be used to improve seed-management recommendations for foresters LQ%UD]LO&KLOHDQGWKH8QLWHG6WDWHVZKRDUHSURGXFLQJVHHGOLQJVRIERWKVSHFLHV in nurseries for ex situ conservation seed orchards as well as for land managers throughout eastern North America interested in growing native hemlocks for refor- estation purposes. 0DWHULDOVDQG0HWKRGV :HFROOHFWHG ULSH VHHG FRQHV LQ 6HSWHPEHU &DUROLQD +HPORFN DQG 2FWREHU (DVWHUQ+HPORFN IURPQDWXUDOVWDQGV SHUVSHFLHV ORFDWHGLQWKHFHQWUDO and southern Appalachian Mountains of the eastern United States (Table 1). We placed seed cones into cloth bags and stored them in a dry greenhouse ven- WLODWHGWRDPELHQWFRQGLWLRQVIRUGWRIDFLOLWDWHWKHRSHQLQJRIFRQHVDQGUHOHDVH RIVHHGVIURPWKHFRQHVFDOHV .DUUIDOW $IWHUFRQHGU\LQJZHH[WUDFWHGVHHGV from the cones using a shaker box, de-winged them in a small tumbling drum, and removed empty seeds, loose wings, and chaff using a seed blower (Seedburo 0RGHO6HHGEXUR(TXLSPHQW&RPSDQ\'HV3ODLQHV,/ :HUHPRYHGPRUH HPSW\VHHGVE\FRQGXFWLQJDÀRDWWHVWLQZDWHU:HGHWHUPLQHGWKHPRLVWXUHFRQWHQW RIDVXEVHWRIVHHGVIURPHDFKVHHGORWXVLQJD0HWWOHU7ROHGRPRLVWXUHDQDO\]HU 0HWWOHU7ROHGR,QF&ROXPEXV2+ DYHUDJHVHHGPRLVWXUHFRQWHQWZDV IRU(DVWHUQ+HPORFNDQGIRU&DUROLQD+HPORFN:HSDFNDJHGWKHUHPDLQLQJ VHHGVLQVPDOOUHVHDODEOHSODVWLFEDJVDQGSODFHGWKHPLQGU\±FROGVWRUDJHDW& IRUGD\VSULRUWRWKHEHJLQQLQJRIWKHVWXG\ 3UHJHUPLQDWLRQ WUHDWPHQWV FRQVLVWHG RI FROG±PRLVW VWUDWL¿FDWLRQ KHUHDIWHU VWUDWL¿FDWLRQ RIVHHGVIRUDQGGD\VDW&LQDGDUNZDON LQFRROHU)RUVWUDWL¿FDWLRQZHVRDNHGDOOVHHGVLQGLVWLOOHGZDWHUIRUKDWURRP WHPSHUDWXUHDQGSODFHGWKHPLQFP3HWULGLVKHVZLWKDVXEVWUDWHRIZKLWHJHUPLQD- WLRQSDSHUPRLVWHQHGWRVDWXUDWLRQZLWKGLVWLOOHGZDWHU(DFK3HWULGLVKFRQWDLQHG VHHGVDQGWKHUHZHUHGLVKHVSHUVSHFLHVWUHDWPHQWSURYHQDQFHFRPELQDWLRQIRUD WRWDORISHWULGLVKHVDQGVHHGVLQWKHVWXG\:HFKHFNHGGLVKHVGDLO\DQG Table 1. Provenance location and Hemlock Woolly Adelgid infestation status for Eastern Hemlock DQG&DUROLQD+HPORFNVHHGVRXUFHVXWLOL]HGLQWKHVHHGVWUDWL¿FDWLRQVWXG\/HDVWVTXDUHPHDQWRWDO JHUPLQDWLRQ 6( DFURVVDOOFROG±PRLVWVWUDWL¿FDWLRQWUHDWPHQWVLVDOVRUHSRUWHGIRUHDFKVHHGVRXUFH
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