现代月季6个品种花粉超低温保存研究
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摘要
【目的】为实现月季花粉长期保存,满足杂交育种和种质资源保存需要,本文以6个现代月季品种的花粉为试验材料,研究其超低温保存技术。【方法】用扫描电子显微镜观察比较6个品种花粉形态、干燥法测定花粉自然含水量、悬滴法检测花粉活力,筛选出适宜萌发培养液,在此基础上进行了液氮保存,并研究这些因素对超低温保存效果的影响。【结果】6个品种花粉悬滴培养的适宜萌发液为15%蔗糖+0.05%硼酸。不同品种新鲜花粉萌发率不同,春季采收的新鲜花粉萌发率为8.91%～27.07%,秋季为9.95%～24.32%,6个品种春季采集的花粉萌发率均高于秋季,但3个差异显著,3个差异不显著。不同品种新鲜花粉含水量不同,春季采收的花粉含水量为9.6%～11.9%;秋季为9.2%～11.5%,4个品种春、秋两季采收的花粉含水量没有显著差异,另外2个品种春季采收的花粉含水量分别显著高于或低于秋季花粉2.4%或1.6%,含水量变化值差异不大。超低温保存后花粉萌发率变化因品种和花粉采收季节而异,6个月季品种花粉经过液氮保存1个月后萌发率为10.36%～21.28%,为新鲜花粉的59.3%～133.9%。长球形的‘金凤凰’和‘希望’,超低温保存后花粉活力有所下降;而超长球形的‘金玛丽’、‘雨果’、‘伦特娜’和‘仙境’,超低温保存后萌发率基本保持稳定。花粉含水量与花粉超低温保存前后萌发率未显著相关。【结论】不同季节采收的自然成熟散粉的6个月季品种花粉均能实现超低温保存,秋季采收的花粉液氮保存后萌发率较新鲜花粉变化较小,但是春季采收的花粉萌发率更高,更适合超低温保存。超低温保存效果与品种、新鲜花粉萌发率、花粉形状和采收季节有关。
【Objective】In order to realize long-term preservation of rose pollen and meet the needs of resource conservation and hybridization breeding, 6 modern rose plants were selected to study the ultra-low temperature preservation technology of their pollen.【Method】The preservation of liquid nitrogen was carried out after the comparison of pollen morphology, pollen viability and water content of six varieties and the selection of suitable germination medium, and the effect of these factors on the cryopreservation was studied.【Result】The results showed that 15 % sucrose + 0.05 % boric acid was suitable for the germination of pollen suspension culture of 6 cultivars. The fresh pollen germination rate of different cultivars was different, it was 8.91 % to 27.07 % in spring, and 9.95 % to 24.32 % in autumn. Six cultivars pollens in spring had a germination rate higher than autumn, but three were significant, and three were not significant. Fresh pollen of different cultivars had different water content which was 9.6 %-11.9 % in spring, and 9.2 %-11.5 % in autumn. The water content of four cultivars pollens harvested in spring and autumn had no significant difference, while the water content of other two cultivars pollens harvested in spring was 2.4 % or 1.6 % respectively higher or lower than it in autumn, showing little significant difference. The changes of pollen germination rate after cryopreservation were different due to the cultivars and harvest season. The germination rate of the six cultivars pollens was 10.36 %-21.28 % after cryopreservation for one month, and it was 59.3 %-133.9 % of the fresh pollen. The pollen feature of‘Golden Scepter',‘Kiboh'was prolate, and the germination rate decreased after cryopreservation.While the pollen feature of‘Goldmarie',‘Victor Hugo',‘Len Turner',‘Carefree Wonder'was perprolate, and the germination rate remained stable. There was no significant correlation between pollen water content and pollen germination rate before and after cryopreservation.【Conclusion】The pollen collected from 6 cultivars and different seasons can be preserved under ultra-low temperature. Although the change of germination rate before and after cryopreservation in autumn was small, the germination rate of pollen harvested in spring was higher, and which was more suitable for cryopreservation. The effect of cryopreservation was related to variety, fresh pollen germination rate, pollen shape and harvest season.
【Objective】In order to realize long-term preservation of rose pollen and meet the needs of resource conservation and hybridization breeding, 6 modern rose plants were selected to study the ultra-low temperature preservation technology of their pollen.【Method】The preservation of liquid nitrogen was carried out after the comparison of pollen morphology, pollen viability and water content of six varieties and the selection of suitable germination medium, and the effect of these factors on the cryopreservation was studied.【Result】The results showed that 15 % sucrose + 0.05 % boric acid was suitable for the germination of pollen suspension culture of 6 cultivars. The fresh pollen germination rate of different cultivars was different, it was 8.91 % to 27.07 % in spring, and 9.95 % to 24.32 % in autumn. Six cultivars pollens in spring had a germination rate higher than autumn, but three were significant, and three were not significant. Fresh pollen of different cultivars had different water content which was 9.6 %-11.9 % in spring, and 9.2 %-11.5 % in autumn. The water content of four cultivars pollens harvested in spring and autumn had no significant difference, while the water content of other two cultivars pollens harvested in spring was 2.4 % or 1.6 % respectively higher or lower than it in autumn, showing little significant difference. The changes of pollen germination rate after cryopreservation were different due to the cultivars and harvest season. The germination rate of the six cultivars pollens was 10.36 %-21.28 % after cryopreservation for one month, and it was 59.3 %-133.9 % of the fresh pollen. The pollen feature of‘Golden Scepter',‘Kiboh'was prolate, and the germination rate decreased after cryopreservation.While the pollen feature of‘Goldmarie',‘Victor Hugo',‘Len Turner',‘Carefree Wonder'was perprolate, and the germination rate remained stable. There was no significant correlation between pollen water content and pollen germination rate before and after cryopreservation.【Conclusion】The pollen collected from 6 cultivars and different seasons can be preserved under ultra-low temperature. Although the change of germination rate before and after cryopreservation in autumn was small, the germination rate of pollen harvested in spring was higher, and which was more suitable for cryopreservation. The effect of cryopreservation was related to variety, fresh pollen germination rate, pollen shape and harvest season.
引文
[1]陈俊愉.中国十大名花[M].上海:上海文化出版社,1989.
[2]Thomas C.Modern roses XI:the world encyclopedia of roses[M].San Diego:Academic Press,2000.
[3]李洪涛,陈海燕,李东伟,等.7个月季品种花粉活力的测定[J].河南农业科学,2014(8):117-119.
[4]赵红霞,王晶,丁晓六,等.蔷薇属植物与现代月季品种杂交亲和性研究[J].西北植物学报,2015,35(4):743-753.
[5]张亚利,尚晓倩,刘燕.花粉超低温保存研究进展[J].北京林业大学学报,2006(4):139-147.
[6]王红,王春梅,黄鲲,等.柑桔花粉超低温保存初探[J].中国南方果树,2006(4):8-10.
[7]石思信,张志娥,肖建平,等.玉米花粉超低温长期保存后遗传稳定性的研究[J].作物学报,1996(4):409-413.
[8]王玉萍,张峰,王蒂,等.马铃薯花粉的超低温保存研究[J].园艺学报,2013,30(6):683-686.
[9]樊新民,牛建新,孙爱新,等.榆叶梅和海棠花粉萌发率的测定及超低温保存方法[J].湖南农业科学,2004(6):65-66.
[10]李广清.山茶花粉超低温保存研究[D].北京林业大学硕士学位论文,2005:1-69.
[11]徐瑾.玉兰花粉超低温保存机制研究[D].北京林业大学博士学位论文,2014:1-171.
[12]孙宪芝.北林月季杂交育种技术体系初探[D].北京林业大学硕士学位论文,2004:1-76.
[13]韩进.月季种质资源创新技术的研究[D].山东农业大学硕士学位论文,2005:1-68.
[14]张立磊,李桂荣,娄腾雪,等.不同处理对“丰花月季”花粉生活力的影响[J].北方园艺,2015(16):72-75.
[15]沈捷,祝晨辰,徐进,等.硼、钙离子对杉木花粉萌发和花粉管生长影响[J].林业工程学报,2010,24(6):57-60.
[16]Meng X J,Shi Y,Wei D,et al.ROS-induced oxidative stress is closely related to pollen deterioration following cryopreservation[J].Cryopreservation,2017,53:433-439.
[17]石印.园林植物花粉超低温保存中的氧化应激研究[D].北京:北京林业大学硕士学位论文,2015:1-86.
[18]张亚利.梅花花粉超低温保存研究及其花粉库建立[D].北京:北京林业大学博士学位论文,2007:1-142.
[19]李秉玲.芍药属植物超低温保存花粉的差异表达蛋白质研究及花粉库的建立[D].北京:北京林业大学博士学位论文,2010:1-168.
[20]Towill L E,Walters C.Cryopreservation of pollen[M].Cryopreservation of tropical germplasm,current research progress and application,Engelmann F,Takagi H,Ibaraki Japan International Research Center for Agricultural Sciences,2000:115-129.
[21]Vendrame W A,Carvalho V S,Dias J M M,et al.Pollination of Dendrobium hybrids using cryopreserved pollen[J].HORTSCIENCE.2008,43(1):264-267.
[22]时婷婷,张桂玲,温四民,等.百合花粉超低温保存方法研究[J].北方园艺,2013(3):64-66.
[23]周守标,余本祺.石蒜属植物花粉形态及分类研究[J].园艺学报,2005,32(5):914-917.
[24]Walker J W.原始被子植物花粉外壁进化的意义[M].中科院植物所古植物研究室孢粉组译.北京:科学出版社,1980.
[25]Wodehouse R P.Pollen Grains[M].New York:Me Graw Hil Book Co.Inc.,1935:323-340.
[2]Thomas C.Modern roses XI:the world encyclopedia of roses[M].San Diego:Academic Press,2000.
[3]李洪涛,陈海燕,李东伟,等.7个月季品种花粉活力的测定[J].河南农业科学,2014(8):117-119.
[4]赵红霞,王晶,丁晓六,等.蔷薇属植物与现代月季品种杂交亲和性研究[J].西北植物学报,2015,35(4):743-753.
[5]张亚利,尚晓倩,刘燕.花粉超低温保存研究进展[J].北京林业大学学报,2006(4):139-147.
[6]王红,王春梅,黄鲲,等.柑桔花粉超低温保存初探[J].中国南方果树,2006(4):8-10.
[7]石思信,张志娥,肖建平,等.玉米花粉超低温长期保存后遗传稳定性的研究[J].作物学报,1996(4):409-413.
[8]王玉萍,张峰,王蒂,等.马铃薯花粉的超低温保存研究[J].园艺学报,2013,30(6):683-686.
[9]樊新民,牛建新,孙爱新,等.榆叶梅和海棠花粉萌发率的测定及超低温保存方法[J].湖南农业科学,2004(6):65-66.
[10]李广清.山茶花粉超低温保存研究[D].北京林业大学硕士学位论文,2005:1-69.
[11]徐瑾.玉兰花粉超低温保存机制研究[D].北京林业大学博士学位论文,2014:1-171.
[12]孙宪芝.北林月季杂交育种技术体系初探[D].北京林业大学硕士学位论文,2004:1-76.
[13]韩进.月季种质资源创新技术的研究[D].山东农业大学硕士学位论文,2005:1-68.
[14]张立磊,李桂荣,娄腾雪,等.不同处理对“丰花月季”花粉生活力的影响[J].北方园艺,2015(16):72-75.
[15]沈捷,祝晨辰,徐进,等.硼、钙离子对杉木花粉萌发和花粉管生长影响[J].林业工程学报,2010,24(6):57-60.
[16]Meng X J,Shi Y,Wei D,et al.ROS-induced oxidative stress is closely related to pollen deterioration following cryopreservation[J].Cryopreservation,2017,53:433-439.
[17]石印.园林植物花粉超低温保存中的氧化应激研究[D].北京:北京林业大学硕士学位论文,2015:1-86.
[18]张亚利.梅花花粉超低温保存研究及其花粉库建立[D].北京:北京林业大学博士学位论文,2007:1-142.
[19]李秉玲.芍药属植物超低温保存花粉的差异表达蛋白质研究及花粉库的建立[D].北京:北京林业大学博士学位论文,2010:1-168.
[20]Towill L E,Walters C.Cryopreservation of pollen[M].Cryopreservation of tropical germplasm,current research progress and application,Engelmann F,Takagi H,Ibaraki Japan International Research Center for Agricultural Sciences,2000:115-129.
[21]Vendrame W A,Carvalho V S,Dias J M M,et al.Pollination of Dendrobium hybrids using cryopreserved pollen[J].HORTSCIENCE.2008,43(1):264-267.
[22]时婷婷,张桂玲,温四民,等.百合花粉超低温保存方法研究[J].北方园艺,2013(3):64-66.
[23]周守标,余本祺.石蒜属植物花粉形态及分类研究[J].园艺学报,2005,32(5):914-917.
[24]Walker J W.原始被子植物花粉外壁进化的意义[M].中科院植物所古植物研究室孢粉组译.北京:科学出版社,1980.
[25]Wodehouse R P.Pollen Grains[M].New York:Me Graw Hil Book Co.Inc.,1935:323-340.
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