香蕉杂交种‘粉杂1号’ABBB基因组鉴定

doi:10.16420/j.issn.0513-353x.2021-0514

摘要/Abstract

摘要：

香蕉杂交品种‘粉杂1号’是当前商业种植中唯一的抗香蕉枯萎病粉蕉（Musa spp. ABB,Pisang awak）品种,该品种来源于高感病的‘广粉1号’（ABB）偶然得到的种子实生苗,其基因组组成和父本未知。通过流式细胞技术（FCM）、染色体制片、基于ITS的限制性片段长度多态性聚合酶链反应（ITS-PCR-RFLP）及反转录转座子间扩增多态性（copia-IRAP）分子标记技术,并结合形态性状评分对‘粉杂1号’基因组组成进行鉴定。流式细胞分析结果显示‘粉杂1号’为四倍体,染色体制片显示染色体数为2n = 4x = 44条,ITS-PCR-RFLP分子标记结果显示其同时具有A和B基因组,copia-IRAP结果显示其含两套或者以上B基因组;形态打分为67分,对应基因组组成为ABBB。综合以上结果认为‘粉杂1号’基因组组成为ABBB,推测其父本可能为BB基因组组成的野生蕉。

关键词: 香蕉, 杂交种, 基因组, 分子标记, 性状评分

Abstract:

The banana cultivar‘Fenza 1’is the only Fenjiao（Musa spp. ABB,Pisang awak）cultivar with resistance to Fusarium wilt in the current commercial banana plantings. The cultivar came from the seeds seedling obtained accidently. Its genomic composition and male parent are unknowen. In this study,Flow cytometry（FCM）,chromosome flaking,molecular marker technology ITS-PCR-RFLP and copia-IRAP,cooperate with characters taxonomic scorecard were used to identify the genomic composition of‘Fenza 1’. The ploidy was tetraploid showed by the FCM. The chromosome number was 2n = 4x = 44 showed by the chromosome flaking;its genomic composition include A and B showed by the ITS-PCR-RFLP;two or more than two B genomic groups showed by the copia-IRAP;the score of the characters taxonomic is 67,and the corresponding genomic composition is ABBB. Based on the results above,we prosposed that the genomic composition of‘Fenza1’is ABBB,and speculated its male parent may be wild banana with the genomic composition BB.

Key words: banana, hybrid, genome, molecular marker, characters taxonomic scorecard

中图分类号:

S668.1

杨兴玉, 许林兵, 吴元立, 邱迪洋, 范琳琳, 黄秉智. 香蕉杂交种‘粉杂1号’ABBB基因组鉴定[J]. 园艺学报, 2022, 49(9): 1991-1997.

YANG Xingyu, XU Linbing, WU Yuanli, QIU Diyang, FAN Linlin, HUANG Bingzhi. Genome（ABBB）Identification of a Hybrid Banana Cultivar‘Fenza 1’[J]. Acta Horticulturae Sinica, 2022, 49(9): 1991-1997.

图/表 5

图1 粉蕉‘粉杂1号’流式细胞荧光测定图 a. 长梗蕉;b.‘广粉1号’;c. 粉杂1号。

Fig. 1 Flow cytometricfluorescent detection images of‘Fenza 1’ a. Musa balbisiana（BBw）;b.‘Guangfen 1’;c.‘Fenza 1’.

图2 ‘粉杂1号’的间期核（a）,前期（b）和中期（c）染色体

Fig. 2 The resting nuclei（a）,prophase（b）and metaphase chromosomes（c）of‘Fenza 1’

图3 ‘粉杂1号’ITS（RsaⅠ,a）和copia-IRAP（AluⅠ,b）引物PCR扩增电泳结果

Fig. 3 Electrophoresis of the PCR amplification of ITS（RsaⅠ,a）and copia-IRAP（AluⅠ,b）from‘Fenza1’

图4 ‘粉杂1号’粉蕉分类性状图 a：假茎;b：叶柄沟槽;c：穗柄;d：果柄;e：胚珠;f：雄花苞片褪色、形状及肩;g：苞片痕、尖、卷起及外表颜色;h：雄花游离花瓣;i：雄花颜色;j：柱头。

Fig. 4 Characters taxonomic illustration of banana‘Fenza 1’ a：Pseudostem;b：Petiole canal;c：Peduncle;d：Fruit pedicel;e：Ovule;f：Bract fading,shape and shoulder,g：Bracts scars,curling and outside color;h：Free petal of male flowers;i：Male flower color;j：Stigma.

表1 ‘粉杂1号’粉蕉性状评分表

Table 1 Characters taxonomic scorecard of banana‘Fenza 1’

性状 Character	粉杂1号Fenza 1	评分score
假茎着色Pseudostem color	无斑点Blotches slight or absent marked	5
叶柄沟槽Petiole canal	边缘闭合下面不具翼膜,抱假茎 Margin inclosed,not winged below,clasping pseudostem	5
穗柄Peduncle	无毛 Glabrous	5
果柄Fruit pedicel	微长 Slightly long	4
胚珠Ovule	每个心室3 ~ 4行胚珠不规则排列 Every four lines within the ovary ovule irregular arrangement	5
苞片肩Bract shoulder	低,比值≈ 0.30 Low,ratio ≈ 0.30	4
苞片卷曲程度Bract curling	苞片抬起,不反卷 Bracts lift but do not roll back	5
苞片形状Bract shape	卵形 Oval	3
苞片顶端Bract apex	钝尖 Obtuse	3
苞片色泽Bract color	外部为棕红色,内部为明亮的深红色 Brownish red outside,bright crimson inside	5
颜色褪色Color fading	苞片内表面颜色连续一致 Inside bract color contimuous to base	5
苞片痕Bract scars	稍微突出 Scarcely prominnent	4
雄花游离花瓣 Free tepal of male flower	顶端下面几乎无褶皱 Rarely corrugated below tip	4
雄花颜色Male flower color	紫红 Pink-Purple	5
柱头颜色Stigma color	粉淡黄色 Pale Yellow	5

参考文献 15

[1]	D'Hont A. 2005. Unraveling the genome structure of polyploids using FISH and GISH;examples of sugarcane and banana. Cytogenetic and GenomeResearch,(109):27-33.
[2]	D'Hont A, Denoeud F, Aury J M, Baurens F C, Carreel F, Garsmeur O, Noel B, Bocs S, Droc G, Rouard M, Da Silva C, Jabbari K, Cardi C, Poulain J, Souquet M, Labadie K, Jourda C, Lengelle J, Rodier-Goud M, Alberti A, Bernard M, Correa M, Ayyampalayam S, McKain M R, Leebens-Mack J, Burgess D, Freeling M, Mbeguie A M D, Chabannes M, Wicker T, Panaud O, Barbosa J, Hribova E, Heslop-Harrison P, Habas R, Rivallan R, Francois P, Poiron C, Kilian A, Burthia D, Jenny C, Bakry F, Brown S, Guignon V, Kema G, Dita M, Waalwijk C, Joseph S, Dievart A, Jaillon O, Leclercq J, Argout X, Lyons E, Almeida A, Jeridi M, Dolezel J, Roux N, Risterucci A M, Weissenbach J, Ruiz M, Glaszmann J C, Quetier F, Yahiaoui N, Wincker P. 2012. The banana(Musa acuminata)genome and the evolution of monocotyledonous plants. Nature, 7410 (488):213-217.
[3]	Huang Bingzhi. 2006. Descriptors and data standard for banana. Beijing: China Agricultural Publishing House. (in Chinese)
	黄秉智. 2006. 香蕉种质资源描述规范和数据标准. 北京: 中国农业出版社.
[4]	Jeridi M, Bakry F, Escoute J, Fondi E, Carreel F, Ferchichi A, D'Hont A, Rodier-Goud M. 2011. Homoeologous chromosome pairing between the A and B genomes of Musa spp. revealed by genomic in situ hybridization. Annals of Botany, 108 (5):975-981. doi: 10.1093/aob/mcr207 pmid: 21835815
[5]	Kuang Ruibin, Li Chunyu, Yang Jing, Wei Yuerong, Yang Qiaosong, Hu Chunhua, Sheng Ou, Yi Ganjun. 2013. Cell structure alteration of banana cultivars with different resistance to Fusarium oxysporum f. Sp. cubense. Molecular Plant Breeding, 11 (2):193-198. (in Chinese)
	邝瑞彬, 李春雨, 杨静, 魏岳荣, 杨乔松, 胡春华, 盛鸥, 易干军. 2013. 抗感枯萎病香蕉的细胞结构抗性研究. 分子植物育种, 11 (2):193-198.
[6]	Li Weiming, Chen Jingjing, Duan Yajie, Pang Zhencai, Sun Dequan, Hu Yulin, Hu Huigang, Xie Jianghui, Chen Wenna. 2018. Research advances on taxonomy,geo-distribution and molecular phylogenetics of wild banana germplasm resources. Acta Horticulturae Sinica, 45 (9):1675-1687. (in Chinese)
	李伟明, 陈晶晶, 段雅婕, 庞振才, 孙德权, 胡玉林, 胡会刚, 谢江辉, 陈文娜. 2020. 香蕉野生种质资源的分类、分布和分子系统发育研究进展. 园艺学报, 45 (9):1675-1687.
[7]	Liu Wenqing, Zhou Jiankun, Li Hongbo, Yang Henian, Lü Shun, Zhuang Huacai. 2010. Introduction evaluation of new banana cultivar resistant to Fusarium wilt disease. Guangdong Agricultural Sciences, 37 (3):110-113. (in Chinese)
	刘文清, 周建坤, 李洪波, 杨贺年, 吕顺, 庄华才. 2010. 抗枯萎病香蕉新品种引种评价. 广东农业科学, 37 (3):110-113.
[8]	Lü Shun, Ren Yi, Wang Fang, Hu Guibing, Huang Bingzhi, Liu Wenqing, He Jianqi, Liu Jianping, Zeng Lisha, Zhou Jiankun, Mai Jingyu, Zhang Keheng. 2018. Ploidy identification of 169 Musa germplasms by flow cytometry. Journal of Fruit Science, 35 (6):668-684. (in Chinese)
	吕顺, 任毅, 王芳, 胡桂兵, 黄秉智, 刘文清, 何建齐, 刘建平, 曾莉莎, 周建坤, 麦景郁, 张珂恒. 2018. 利用流式细胞术快速鉴定169份香蕉种质资源的染色体倍性. 果树学报, 35 (6):668-684.
[9]	Martin K P, Pachathundikandi S K, Zhang C L, Slater A, Madassery J H. 2006. RAPD analysis of a variant of banana(Musa sp.) cv. grande naine and its propagation via shoot tip culture. In Vitro Cellular & Developmental Biology - Plant, 42 (2):188-192.
[10]	Nair A S, Teo C H, Schwarzacher T, Harrison P H. 2005. Genome classification of banana cultivars from South India using IRAP markers. Euphytica, 144 (3):285-290. doi: 10.1007/s10681-005-7321-2 URL
[11]	Nwakanma D C, Pillay M, Okoli B E, Tenkouano A. 2003. PCR-RFLP of the ribosomal DNA internal transcribed spacers(ITS) provides markers for the A and B genomes in Musa L. Theoretical and Applied Genetics, 108 (1):154-159 pmid: 12955208
[12]	Simmonds N W, Shepherd K. 1955. The taxonomy and origins of the cultivated bananas. Journal of the Linnean Society of London,Botany, 359 (55):302-312.
[13]	Simmonds N W, Weatherup S T C. 1990. Numerical taxonomy of the wild bananas(Musa). New Phytologist, 115 (3):567-571 doi: 10.1111/j.1469-8137.1990.tb00485.x pmid: 33874268
[14]	Wang Fang, Niu Yuqing, Huang Bingzhi, Cui Guangjuan, Zhang Keheng, Zeng Lisha, Liu Wenqing, Lü Shun, Guo Quanxiang, He Jianqi. 2019. Studies on SCAR marker related with B genome in Musa. Acta Horticulturae Sinica, 46 (3):577-589. (in Chinese)
	王芳, 牛玉清, 黄秉智, 崔广娟, 张珂恒, 曾莉莎, 刘文清, 吕顺, 郭权香, 何建齐. 2019. 香蕉与B基因组相关的SCAR标记研究. 园艺学报, 46 (3):577-589. doi: 10.16420/j.issn.0513-353x.2018-0430 URL
[15]	Wei Huafang, Li Chaosheng, Wei Shaolong, Wei Di, Zhang Jinzhong, Wei Zhiheng. 2012. Preliminary report on the introduction test of Musa AAB Group Fenjiao variety‘Fenza 1’in Guangxi. Journal of Southern Agriculture, 43 (9):1353-1356. (in Chinese)
	韦华芳, 李朝生, 韦绍龙, 韦弟, 张进忠, 韦志横. 2012. “粉杂1号”粉蕉广西引种试验初报. 南方农业学报, 43 (9):1353-1356.