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园艺学报 ›› 2020, Vol. 47 ›› Issue (12): 2451-2458.doi: 10.16420/j.issn.0513-353x.2020-0769

• 新技术与新方法 • 上一篇    下一篇

依据多倍体形态特征快速高效发掘柑橘四倍体

周 锐1,解凯东1,王 伟1,彭 珺1,谢善鹏1,胡益波2,伍小萌1,郭文武1,*   

  1. 1华中农业大学园艺林学学院,园艺植物生物学教育部重点实验室,武汉 430070;2肇庆市德庆县柑桔研究所,广东肇庆 526600
  • 出版日期:2020-12-25 发布日期:2021-01-06
  • 基金资助:
    国家重点研发计划项目(2019YFD1000100);国家自然科学基金项目(31820103011,31672108);广东省科技计划项目(2018B020202009);湖北省科技计划项目(2020BBA036);中央高校基本科研业务费专项资金资助项目(2662019QD048)

Efficient Identification of Tetraploid Plants from Seedling Populations of Apomictic Citrus Genotypes Based on Morphological Characteristics

ZHOU Rui1,XIE Kaidong1,WANG Wei1,PENG Jun1,XIE Shanpeng1,HU Yibo2,WU Xiaomeng1,and GUO Wenwu1,*   

  1. 1Key laboratory of Horticultural Plant Biology(Ministry of Education),College of Horticulture and Forestry Sciences,Huazhong Agricultural University,Wuhan 430070,China;2Citrus Research Institute of Deqing County,Zhaoqing,Guangdong 526600,China
  • Online:2020-12-25 Published:2021-01-06

摘要: 基于柑橘多胚性品种存在珠心细胞自然加倍的特点,建立了基于种子催芽、生长室实生播种、幼苗“观根辨叶看油胞”形态初选和流式细胞仪倍性鉴定的发掘柑橘四倍体的快速方法。采用该方法,分别从无酸甜橙、红江橙、贡柑、年橘、新会柑、滑皮金柑的432、506、1 356、1 870、2 749和685株实生苗中,发掘出疑似四倍体5、6、5、17、45、7株;通过倍性检测,从上述6个品种分别获得四倍体4、5、5、14、39、5株,形态初选准确率分别为80.0%、83.3%、100.0%、82.4%、86.7%和71.4%。与之前的四倍体发掘方法相比,本方法四倍体初选准确率高,并且从种子播种到获得四倍体植株耗时不超过40 d。SSR分子鉴定表明,所鉴定的四倍体均来自其二倍体品种珠心细胞自然加倍形成的双二倍体,为柑橘三倍体无核育种提供了珍贵的四倍体亲本资源。

关键词: 柑橘, 多倍体, 流式细胞仪, SSR分子标记

Abstract: Based on the phenomenon of spontaneous doubling of nucellar cells in polyembryonic citrus genotypes,we established a simple and efficient approach for citrus tetraploid exploitation by sowing the seeds(after germination being accelerated in a 28 ℃ incubator for about one week)in pots in a controlled growth chamber,followed by screening tetraploids from germinated seedlings based on root,leaf morphology,oil gland density and flow cytometry analysis. By this method,we screened 5,6,5,17,45 and 7 putative tetraploids respectively from the 432,506,1 356,1 870,2 749 and 685 seedlings of Succari sweet orange,Hongjiang sweet orange,Gonggan mandarin,Nian mandarin,Xinhui mandarin and Huapi kumquat. By analyzing their ploidy level using flow cytometry,a total of 72 tetraploids was obtained,including 4 from Succari sweet orange,5 from Gonggan mandarin,5 from Hongjiang sweet orange,14 from Nian mandarin,39 from Xinhui mandarin and 5 from Huapi kumquat,with the tetraploid pre-screening accuracy rate of 80.0%,83.3%,100.0%,82.4%,86.7% and 71.4%,respectively. The merits of our new method are higher accuracy rate of tetraploid pre-screening,and time saving only needing less than 40 d from seed sowing to tetraploid confirmation. Simple sequence repeat(SSR)analysis showed that the bands of all tetraploids obtained herein are identical with that of their corresponding diploid genotypes,indicating that all these plants are the doubled diploids which have great potential for seedless triploid breeding.

Key words: citrus, polyploid, flow cytometry, SSR molecular marker

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