二倍体香蕉种质资源花粉活力检测方法的筛选  

Selection of Testing Methods on Pollen Viability of Diploid Banana(Musa spp.)Accessions

作　　者：邓彪[1,2] 盛鸥[2] 魏岳荣[2] 李大志[1] 高慧君[2] 邝瑞彬[2] 易干军[2] Agustin B.Molina 

机构地区：[1]湖南农业大学园艺园林学院,长沙410128 [2]农业部南亚热带果树生物学与遗传资源利用重点实验室、广东省农业科学院果树研究所,广州510640 [3]国际生物多样性组织-亚太香蕉协作网,洛斯班尼奥斯4031

出　　处：《分子植物育种》2014年第5期1011-1017,共7页Molecular Plant Breeding

基　　金：国家自然科学基金项目(U1131004;31101510);863计划项目(2011AA10020603);科技部国际合作专项(2013DFB30400);948计划项目(2011-G16);国家香蕉产业技术体系建设项目(CARS-32-01);广东省现代农业产业技术体系专项(lnsgtx-03);广州市“珠江科技新星”专项(2013J2200081);Bioversity International项目(LoA2012/48)共同资助

摘　　要：本研究利用4种父本育性较强的二倍体香蕉(Musa spp.)种质,比较不同染色法和离体萌发法检测香蕉花粉活力的效果,以期筛选适合香蕉花粉活力快速检测的方法。结果显示:醋酸洋红和I-KI染色法处理后的香蕉花粉形态较模糊,而TTC法与MTT法的染色效果较好,花粉形态较清晰,容易区分染色与未染色的花粉;适合香蕉花粉离体萌发的培养基中,蔗糖浓度为15%,pH值为5.8;MTT染色法结果与离体萌发法结果相关性最好,说明MTT染色法的结果较其他染色法准确、可靠。利用MTT染色法,本研究比较了17份二倍体香蕉种质的花粉活力,结果显示:‘Calcutta 4’(M.acuminata ssp.burmannicoides(AA))、BGY3(M.balbisiana(BB))、Malaccensis(M.acuminata ssp.malaccensis(AA))、Balbisiana(M.balbisiana(BB))等4份野生类型种质,以及双单倍体种质‘CIRAD 930’(M.acuminata ssp.malaccensis var.DH Pahang(AA))的染色率超过50%,而3份改良二倍体种质和4份栽培类型种质的染色率低于30%。We compared staining methods and in vitro germination assays for evalutating banana （Musa spp.） pollen viability, using the four diploids as samples that are known to have high levels of male fertility, aiming to select a quick and effective protocol for determining pollen viability in banana germplasm. The results showed that, most of the pollen grains were causally obscure after stained with acetocarmine and iodine potassium iodide （I-KI）, whereas the stained or un-stained pollen grains were clear to be distinguished when treated with 2, 3, 5 triphenyl tetrazolium chloride （TTC） and 2, 5 diphenyl tetrazolium bromide （MTT）. And for in vitro germination of pollen grains, the optimal sucrose concentration and pH in culture medium were 15% and 5.8, respectively. It was further found that, the results from MTT staining method were well-correlated with the results obtained from that optimal assay of in vitro germination, indicating that MTT staining method might be a more effective protocol for determining pollen viability in banana, compared with other staining methods. Using such selected MTT staining method, we further examined the pollen viability of 17 diploid banana accessions. The results showed that, the mean pollen stainability higher than 50% was found in four wild accessions, ＇Calcutta 4＇ （M. acuminata ssp. burmannicoides （AA））,BGY3 （M. balbisiana （BB））,Malaccensis （M. aeuminata ssp. malaecensis （AA）） and Balbi- siana （M. balbisiana （BB））, and in a double hapolid accession, ＇CIRAD 930＇ （M. acuminata ssp. malaecensis var. DH Pahang （AA））. Whereas, in the three imporved diploids and four cultivated varieties, their pollen stainability were lower than 30%.

关 键 词：二倍体香蕉 花粉生活力 测定方法 MTT染色法 

分 类 号：S668.1[农业科学—果树学]