高温诱导大孢子染色体加倍选育毛白杨杂种三倍体
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摘要
【目的】鉴于杨树三倍体具有突出的营养生长优势,针对毛白杨良种选育中母本效应难以充分利用问题,选择毛白杨育性好的雌株诱导大孢子染色体加倍选育白杨杂种三倍体,为毛白杨三倍体育种提供技术支撑。【方法】本研究以毛白杨无性系MC1和MC2为母本,银腺杨YX1为父本。根据白杨大小孢子发生发育存在相关性的原理,以雄株小孢子发育进程为即时判别大孢子发生进程的参照,当小孢子发育至单核早期、单核靠边期和双核期时,分别施加38、40和42℃高温持续处理雌花序4 h。【结果】两个杂交组合的处理组共得到189株毛白杨杂种三倍体,其中MC1×YX1组合中共获得70株杂种三倍体,平均诱导率为56.45%; MC2×YX1组合中共获得119株杂种三倍体,平均诱导率为47.04%,对照组未获得杂种三倍体。当小孢子发育至单核靠边期,对应大孢子母细胞减数分裂至粗线期至中期Ⅰ,此时施加38和40℃高温处理4 h是毛白杨大孢子染色体加倍的最佳处理时期。两个杂交组合的杂种三倍体在平均苗高和地径生长方面均优于二倍体,但组合间存在一定差异,表明三倍体育种同样需要重视亲本选配。【结论】研究进一步证明高温诱导大孢子染色体加倍是一种高效获得毛白杨杂种三倍体的技术途径,相关技术进步对于推进毛白杨良种选育进程具有重要的意义。
[Objective]Triploid poplars have been proved to be characterized with outstanding advantages in vegetative growth over diploids.However,it still has some difficulties to take full advantage of maternal effect in present P.tomentosa breeding work.By careful selection of female parents with good fertility,this study aims to produce hybrid triploids by megaspore chromosome doubling in P.tomentosa,which would provide technical support for triploid breeding in further research.[Method]The P.tomontosa clones MC1 and MC2 were served as the female parents,and the hybrid clone YX1 of P.alba ×P.glandulosa was selected to be the male parent.According to the correlation between megasporocytes and microsporocytes during meiosis,the meiotic stage of megaspore in female flowers could be determined by the meiotic stage of microspore in male flowers.When the microspore developed into early stage of single nucleus,late uninucleate stage and binucleate stage,the female catkins were treated for 4 hours with high temperatures of 38,40 and 42 ℃ to induce chromosome doubling of megaspore,respectively.[Result]A total of 189 triploids were obtained in the two hybrid combinations.Among all the 189 triploids,70 triploids were obtained from the cross combination MC1×YX1,and the average induction rate was 56.45%.119 triploids were obtained from the combination MC2×YX1,and the mean induction rate was 47.04%.No triploids were achieved in the control groups.When the microspores developed into the uninucleate eccentric stage,the corresponding meiotic stage of macrospore mother cell was at the stages from pachytene to metaphase I,which could be the optimal treating period for inducing megaspore chromosome doubling by 38 and 40 ℃ high temperature treatment.The average seedling height and the ground diameter of triploid hybrids derived from two cross combinations were higher than that of the diploids,amid there were some differences between the two combinations,which suggested that some attentions should be paid to the parental selection in triploid breeding of P.tomentosa.[Conclusion]Our findings reveal that induction of megaspore chromosome doubling by high temperature treatment is an effective way to produce triploid hybrids in P.tomentosa,and the progress of breeding technologies plays an important role in genetic improvement of P.tomentosa.
[Objective]Triploid poplars have been proved to be characterized with outstanding advantages in vegetative growth over diploids.However,it still has some difficulties to take full advantage of maternal effect in present P.tomentosa breeding work.By careful selection of female parents with good fertility,this study aims to produce hybrid triploids by megaspore chromosome doubling in P.tomentosa,which would provide technical support for triploid breeding in further research.[Method]The P.tomontosa clones MC1 and MC2 were served as the female parents,and the hybrid clone YX1 of P.alba ×P.glandulosa was selected to be the male parent.According to the correlation between megasporocytes and microsporocytes during meiosis,the meiotic stage of megaspore in female flowers could be determined by the meiotic stage of microspore in male flowers.When the microspore developed into early stage of single nucleus,late uninucleate stage and binucleate stage,the female catkins were treated for 4 hours with high temperatures of 38,40 and 42 ℃ to induce chromosome doubling of megaspore,respectively.[Result]A total of 189 triploids were obtained in the two hybrid combinations.Among all the 189 triploids,70 triploids were obtained from the cross combination MC1×YX1,and the average induction rate was 56.45%.119 triploids were obtained from the combination MC2×YX1,and the mean induction rate was 47.04%.No triploids were achieved in the control groups.When the microspores developed into the uninucleate eccentric stage,the corresponding meiotic stage of macrospore mother cell was at the stages from pachytene to metaphase I,which could be the optimal treating period for inducing megaspore chromosome doubling by 38 and 40 ℃ high temperature treatment.The average seedling height and the ground diameter of triploid hybrids derived from two cross combinations were higher than that of the diploids,amid there were some differences between the two combinations,which suggested that some attentions should be paid to the parental selection in triploid breeding of P.tomentosa.[Conclusion]Our findings reveal that induction of megaspore chromosome doubling by high temperature treatment is an effective way to produce triploid hybrids in P.tomentosa,and the progress of breeding technologies plays an important role in genetic improvement of P.tomentosa.
引文
[1]康向阳.毛白杨细胞遗传与三倍体选育[D].北京:北京林业大学,1996.Kang X Y.Study on cytogenetic and allotriploid breeding of Populus tomentosa[D].Beijing:Beijing Forestry University,1996.
[2]叶培忠.白杨繁殖育种[J].林业科学,1955,1(1):37--46.Ye P Z.Poplar breeding[J].Scientia Silvae Sinicae,1995,1(1):37-46.
[3]朱之悌.全国毛白杨优树资源收集、保存和利用的研究[J].北京林业大学学报,1992,14(增刊3):1--25.Zhu Z T.Collection,conservation and utilization of plus tree resources of Populus tomentosa in China[J].Journal of Beijing Forestry University,1992,14(Suppl.3):1--25.
[4]林惠斌,朱之悌.毛白杨杂交育种战略的研究[J].北京林业大学学报,1988,10(3):97--101.Lin H B,Zhu Z T.Studies on breeding strategies of Populus tomentosa[J].Journal of Beijing Forestry University,1988,10(3):97-101.
[5]朱之悌,林惠斌,康向阳.毛白杨异源三倍体B301等无性系选育的研究[J].林业科学,1995,31(6):499-505.Zhu Z T,Lin H B,Kang X Y.Studies on allotriploid breeding of Populus tomentosa B301 clones[J].Scientia Silvae Sinicae,1995,31(6):499-505.
[6]张志毅,李凤兰.白杨染色体加倍技术研究及三倍体育种(Ⅰ):花粉染色体加倍技术[J].北京林业大学学报,1992,14(增刊3):52--58.Zhang Z Y,Li F L.Studies on chromosome doubling and triploid breeding of white poplar(Ⅰ):the techniques of the pollen chromosome doubling[J].Journal of Beijing Forestry University,1992,14(Suppl.3):52-58.
[7]康向阳,朱之悌,林惠斌.杨树花粉染色体加倍有效处理时期的研究[J].林业科学,1999,35(4):21-4.Kang X Y,Zhu Z T,Lin H B.Study on the effective treating period for pollen chromosome doubling of Populus tomentosa×P.bolleana[J].Scientia Silvae Sinicae,1999,35(4):21-24.
[8]康向阳,朱之悌,张志毅.高温诱导白杨2n花粉有效处理时期的研究[J].北京林业大学学报,2000,22(3):1--4.Kang X Y,Zhu Z T,Zhang Z Y.Suitable period of high temperature treatment for 2n pollen of Populus tomentosa×P.bolleana[J].Journal of Beijing Forestry University,2000,22(3):1-4.
[9]李云,朱之悌,田砚亭,等.极端温度处理白杨雌花芽培育三倍体植株的研究[J].北京林业大学学报,2000,22(5):7-12.Li Y,Zhu Z T,Tian Y T,et al.Obtaining triploids by high and low temperature treating female flower buds of white poplar[J].Journal of Beijing Forestry University,2000,22(5):7-12.
[10]康向阳,朱之梯,张志毅.银腺杨与毛新杨正反交三倍体选育[J].北京林业大学学报,2000,22(6):8--11.Kang X Y,Zhu Z T,Zhang Z Y.Breeding of triploids by the reciprocal crossing of Populus alba×P.glandulosa and P.tomentosa×P.bolleana[J].Journal of Beijing Forestry University,2000,22(6):8--11.
[11]康向阳,张平冬,高鹏,等.秋水仙碱诱导白杨三倍体新途径的发现[J].北京林业大学学报,2004,26(1):1--4.Kang X Y,Zhang P D,Gao P,et al.Discovery of a new way of poplar triploids induced with colchicine after pollination[J].Journal of Beijing Forestry University,2004,26(1):1-4.
[12]李艳华,康向阳.诱导大孢子染色体加倍选育白杨杂种三倍体[J].北京林业大学学报,2007,29(5):22--25.Li Y H,Kang X Y.Triploid induction in white poplar by chromosome doubling of megaspore[J].Journal of Beijing Forestry University,2007,29(5):22--25.
[13]康宁,白凤莹,张平冬,等.高温诱导胚囊染色体加倍获得毛白杨杂种三倍体[J].北京林业大学学报,2015,37(2):79--86.Kang N,Bai F Y,Zhang P D,et al.Inducing chromosome doubling of embryo sac in Populus tomentosa with high temperature exposure for hybrid triploids[J].Journal of Beijing Forestry University,2015,37(2):79--86.
[14]康向阳,王君.杨树多倍体诱导技术研究[M].北京:科学出版社,2010.Kang X Y,Wang J.Poplar polyploid induction technology research[M].Beijing:Science Press,2010.
[15]张妍,崔彬彬,魏俊杰,等.毛白杨大、小孢子母细胞减数分裂进程的相关性[J].西北农林科技大学学报(自然科学版),2015,43(11):63--70.Zhang Y,Cui B B,Wei J J,et al.Correlation between megesporecytes and microsporecytes meiosis stage of Populus tomentosa Carr.[J].Journal of Northwest A&F University(Nat.Sci.Ed.),2015,43(11):63--70.
[16]Bai F Y,Kang N,Zhang P D,et al.Selection of female parents with high fertility and high combining abilities for cross-breeding Populus tomentosa[J/OL].Journal of Forest Research,2018[2018-07--05].https://link.springer.com/content/pdf/10.1007%2Fs11676-018-0684-8.pdf.
[17]Galbraith D W,Harkins K R,Maddox J M,et al.Rapid flow cytometric analysis of the cell cycle in intact plant tissues[J].Science,1983,220:1049--1051.
[18]Wang J,Li D L,Kang X Y.Induction of unreduced megaspores with high temperature during megasporogenesis in Populus[J].Annals of Forest Science,2011,69(1):59--67.
[19]Lu M,Zhang P D,Kang X Y.Induction of 2n female gametes in Populus adenopoda Maxim by high temperature exposure during female gametophyte development[J].Breed Science,2013,63(1):96--103.
[20]康向阳.关于杨树多倍体育种的几点认识[J].北京林业大学学报,2010,32(5):149-153.Kang X Y.Some understanding on polyploid breeding of poplars[J].Journal of Beijing Forestry University,2010,32(5):149-153.
[21]Ramanna M S,Jacobsen E.Relevance of sexual polyploidization for crop improvement:a review[J].Euphytica,2003,133(1):3-8.
[22]王君.青杨派树种多倍体诱导技术研究[D].北京:北京林业大学,2009.Wang J.Techniques of polyploid induction in Populus spp.(Section Tacamahaca)[D].Beijing:Beijing Forestry University,2009.
[23]Wahid A,Gelani S,Ashraf M,et al.Heat tolerance in plants:an overview[J].Environmental&Experimental Botany,2007,61(3):199-223.
[24]Einspahr D W.Production and utilization of triploid hybrid aspen[J].Iowa State J Res,1984,58(4):401--409.
[25]Weisgerber H,Rau H M,Gartner E J,et al.25 years of forest tree breeding in Hesse[J].Allge Forstze,1980,26:665-712.
[26]Liao T,Cheng S,Zhu X,et al.Effects of triploid status on growth,photosynthesis,and leaf area in Populus[J].Trees,2016,30(4):1137--1147.
[2]叶培忠.白杨繁殖育种[J].林业科学,1955,1(1):37--46.Ye P Z.Poplar breeding[J].Scientia Silvae Sinicae,1995,1(1):37-46.
[3]朱之悌.全国毛白杨优树资源收集、保存和利用的研究[J].北京林业大学学报,1992,14(增刊3):1--25.Zhu Z T.Collection,conservation and utilization of plus tree resources of Populus tomentosa in China[J].Journal of Beijing Forestry University,1992,14(Suppl.3):1--25.
[4]林惠斌,朱之悌.毛白杨杂交育种战略的研究[J].北京林业大学学报,1988,10(3):97--101.Lin H B,Zhu Z T.Studies on breeding strategies of Populus tomentosa[J].Journal of Beijing Forestry University,1988,10(3):97-101.
[5]朱之悌,林惠斌,康向阳.毛白杨异源三倍体B301等无性系选育的研究[J].林业科学,1995,31(6):499-505.Zhu Z T,Lin H B,Kang X Y.Studies on allotriploid breeding of Populus tomentosa B301 clones[J].Scientia Silvae Sinicae,1995,31(6):499-505.
[6]张志毅,李凤兰.白杨染色体加倍技术研究及三倍体育种(Ⅰ):花粉染色体加倍技术[J].北京林业大学学报,1992,14(增刊3):52--58.Zhang Z Y,Li F L.Studies on chromosome doubling and triploid breeding of white poplar(Ⅰ):the techniques of the pollen chromosome doubling[J].Journal of Beijing Forestry University,1992,14(Suppl.3):52-58.
[7]康向阳,朱之悌,林惠斌.杨树花粉染色体加倍有效处理时期的研究[J].林业科学,1999,35(4):21-4.Kang X Y,Zhu Z T,Lin H B.Study on the effective treating period for pollen chromosome doubling of Populus tomentosa×P.bolleana[J].Scientia Silvae Sinicae,1999,35(4):21-24.
[8]康向阳,朱之悌,张志毅.高温诱导白杨2n花粉有效处理时期的研究[J].北京林业大学学报,2000,22(3):1--4.Kang X Y,Zhu Z T,Zhang Z Y.Suitable period of high temperature treatment for 2n pollen of Populus tomentosa×P.bolleana[J].Journal of Beijing Forestry University,2000,22(3):1-4.
[9]李云,朱之悌,田砚亭,等.极端温度处理白杨雌花芽培育三倍体植株的研究[J].北京林业大学学报,2000,22(5):7-12.Li Y,Zhu Z T,Tian Y T,et al.Obtaining triploids by high and low temperature treating female flower buds of white poplar[J].Journal of Beijing Forestry University,2000,22(5):7-12.
[10]康向阳,朱之梯,张志毅.银腺杨与毛新杨正反交三倍体选育[J].北京林业大学学报,2000,22(6):8--11.Kang X Y,Zhu Z T,Zhang Z Y.Breeding of triploids by the reciprocal crossing of Populus alba×P.glandulosa and P.tomentosa×P.bolleana[J].Journal of Beijing Forestry University,2000,22(6):8--11.
[11]康向阳,张平冬,高鹏,等.秋水仙碱诱导白杨三倍体新途径的发现[J].北京林业大学学报,2004,26(1):1--4.Kang X Y,Zhang P D,Gao P,et al.Discovery of a new way of poplar triploids induced with colchicine after pollination[J].Journal of Beijing Forestry University,2004,26(1):1-4.
[12]李艳华,康向阳.诱导大孢子染色体加倍选育白杨杂种三倍体[J].北京林业大学学报,2007,29(5):22--25.Li Y H,Kang X Y.Triploid induction in white poplar by chromosome doubling of megaspore[J].Journal of Beijing Forestry University,2007,29(5):22--25.
[13]康宁,白凤莹,张平冬,等.高温诱导胚囊染色体加倍获得毛白杨杂种三倍体[J].北京林业大学学报,2015,37(2):79--86.Kang N,Bai F Y,Zhang P D,et al.Inducing chromosome doubling of embryo sac in Populus tomentosa with high temperature exposure for hybrid triploids[J].Journal of Beijing Forestry University,2015,37(2):79--86.
[14]康向阳,王君.杨树多倍体诱导技术研究[M].北京:科学出版社,2010.Kang X Y,Wang J.Poplar polyploid induction technology research[M].Beijing:Science Press,2010.
[15]张妍,崔彬彬,魏俊杰,等.毛白杨大、小孢子母细胞减数分裂进程的相关性[J].西北农林科技大学学报(自然科学版),2015,43(11):63--70.Zhang Y,Cui B B,Wei J J,et al.Correlation between megesporecytes and microsporecytes meiosis stage of Populus tomentosa Carr.[J].Journal of Northwest A&F University(Nat.Sci.Ed.),2015,43(11):63--70.
[16]Bai F Y,Kang N,Zhang P D,et al.Selection of female parents with high fertility and high combining abilities for cross-breeding Populus tomentosa[J/OL].Journal of Forest Research,2018[2018-07--05].https://link.springer.com/content/pdf/10.1007%2Fs11676-018-0684-8.pdf.
[17]Galbraith D W,Harkins K R,Maddox J M,et al.Rapid flow cytometric analysis of the cell cycle in intact plant tissues[J].Science,1983,220:1049--1051.
[18]Wang J,Li D L,Kang X Y.Induction of unreduced megaspores with high temperature during megasporogenesis in Populus[J].Annals of Forest Science,2011,69(1):59--67.
[19]Lu M,Zhang P D,Kang X Y.Induction of 2n female gametes in Populus adenopoda Maxim by high temperature exposure during female gametophyte development[J].Breed Science,2013,63(1):96--103.
[20]康向阳.关于杨树多倍体育种的几点认识[J].北京林业大学学报,2010,32(5):149-153.Kang X Y.Some understanding on polyploid breeding of poplars[J].Journal of Beijing Forestry University,2010,32(5):149-153.
[21]Ramanna M S,Jacobsen E.Relevance of sexual polyploidization for crop improvement:a review[J].Euphytica,2003,133(1):3-8.
[22]王君.青杨派树种多倍体诱导技术研究[D].北京:北京林业大学,2009.Wang J.Techniques of polyploid induction in Populus spp.(Section Tacamahaca)[D].Beijing:Beijing Forestry University,2009.
[23]Wahid A,Gelani S,Ashraf M,et al.Heat tolerance in plants:an overview[J].Environmental&Experimental Botany,2007,61(3):199-223.
[24]Einspahr D W.Production and utilization of triploid hybrid aspen[J].Iowa State J Res,1984,58(4):401--409.
[25]Weisgerber H,Rau H M,Gartner E J,et al.25 years of forest tree breeding in Hesse[J].Allge Forstze,1980,26:665-712.
[26]Liao T,Cheng S,Zhu X,et al.Effects of triploid status on growth,photosynthesis,and leaf area in Populus[J].Trees,2016,30(4):1137--1147.