[1]孙博文,黄耀,孙亚亭,等.远缘杂交结合胚培养法获得厚皮甜瓜阿鲁斯达令双单倍体[J].江苏农业学报,2022,38(03):775-781.[doi:doi:10.3969/j.issn.1000-4440.2022.03.024]
　SUN Bo-wen,HUANG Yao,SUN Ya-ting,et al.Production of doubled haploid in muskmelon Earl’s Darling by interspecific hybridization and embryo culture[J].,2022,38(03):775-781.[doi:doi:10.3969/j.issn.1000-4440.2022.03.024]
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远缘杂交结合胚培养法获得厚皮甜瓜阿鲁斯达令双单倍体()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
38
期数:
2022年03期
页码:
775-781
栏目:
园艺
出版日期:
2022-06-30

文章信息/Info

Title:
Production of doubled haploid in muskmelon Earl’s Darling by interspecific hybridization and embryo culture
作者:
孙博文1; 黄耀1; 2; 孙亚亭1; 刘麒1; 2; 郑莉娜1; 孙沛1; 钱春桃1
(1.南京农业大学园艺学院/作物遗传与种质创新国家重点实验室，江苏南京210095；2.南京农业大学新农村发展研究院，江苏苏州215500）
Author(s):
SUN Bo-wen1; HUANG Yao1; 2; SUN Ya-ting1; LIU Qi1; 2; ZHENG Li-na1; SUN Pei1; QIAN Chun-tao1
(1.College of Horticulture, Nanjing Agricultural University/State Key Laboratory of Crop Genetics and Germplasm Innovation, Nanjing 210095, China;2.New Rural Development Research Institute of Nanjing Agricultural University, Suzhou 215500, China)
关键词:
厚皮甜瓜; 远缘杂交; 胚培养; 双单倍体
Keywords:
muskmelon; interspecific hybridization; embryo culture; doubled haploid
分类号:
S652.036
DOI:
doi:10.3969/j.issn.1000-4440.2022.03.024
文献标志码:
A
摘要:
旨在系统地研究通过远缘杂交结合胚培养获得甜瓜双单倍体的方法，以拓宽甜瓜双单倍体获得路径。以西印度瓜（Cucumis anguria L.）为远缘花粉供体，对厚皮甜瓜阿鲁斯达令进行授粉，并结合胚培养方法进行研究，以期获得厚皮甜瓜阿鲁斯达令的双单倍体。结果表明，对阿鲁斯达令甜瓜进行远缘授粉后如果不作处理，子房会全部败育，而对子房表面涂抹坐果灵后坐果率可达73.33%；授粉28 d后，共采收22个远缘杂交果实，累计获得359粒种子；胚培养后获得4株再生植株，再生植株占比为1.11%；通过流式细胞仪鉴定得出，再生植株有二倍体、混倍体2种类型。对比二倍体再生植株和母本阿鲁斯达令发现，二倍体再生苗的叶、花较小，植株长势较弱，果实较圆，平均花粉活力较低。对二倍体再生植株R3、母本阿鲁斯达令自交1代(S1代)群体形态学性状进行遗传变异系数比较发现，R3 S1代群体各项指标的遗传变异系数均远低于母本阿鲁斯达令S1代群体。利用筛选出的2对在阿鲁斯达令基因组中具有多态性的简单重复序列（SSR）引物CMATN240、CMCTN71对R3 S1代群体进行基因型分析，结果表明，20株R3 S1代群体均为纯合带型。通过形态学鉴定、SSR分子标记鉴定分析发现，获得的再生植株R3为双单倍体。由研究结果可知，通过远缘杂交结合胚培养获得了甜瓜再生植株，通过细胞流式仪倍性鉴定、SSR标记分析与S1代群体性状遗传变异系数鉴定得出，再生植株为双单倍体。
Abstract:
This article systematically studied the method of obtaining doubled haploid of melon through interspecific hybridization and embryo culture to broaden the way of obtaining doubled haploids. In this study, Cucumis anguria L. was used as a distant pollen donor to pollinate the thick-skinned muskmelon Earl’s Darling, and embryo culture method was combined to obtain doubled haploids. The results showed that all ovaries were aborted after distant pollination, and the fruit setting rate could reach 73.33% after applying 4-chlorophenoxyacetic acid. After 28 days of pollination, 22 distant hybrid fruits were harvested, and 359 seeds were obtained. Four regenerated plants were obtained after embryo culture, and the proportion of regenerated plants was 1.11%. Flow cytometry identified two types of regenerated plants, diploid and mixoploid. Compared with the female parent, the leaves and flowers of diploid regenerated seedlings were smaller, plant growth was weaker, fruits were rounder, and average pollen vigor was lower. The genetic variation coefficients of the S1 generation of the diploid regenerated plant R3 and the female parent Earl’s Darling were compared, and it was found that the genetic variation coefficients of the indicators of the R3 S1 population were far lower than those of the female parent Earl’s Darling S1 population. Two pairs of polymorphic simple sequence repeat (SSR) primers CMATN240 and CMCTN71 were selected to perform genotype analysis of the R3 S1 population, the results showed that the 20 R3 S1 populations were all homozygous. Morphological identification and SSR molecular marker identification analysis showed that the regenerated plant R3 obtained in this study was a double haploid. The regenerated plants of muskmelon were obtained by distant hybridization and embryo culture. The results of flow cytometry, SSR marker analysis and genetic variation coefficient of S1 population traits show that the regenerated plants are doubled haploids.

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备注/Memo

备注/Memo:
收稿日期：2021-08-30基金项目：江苏省农业重大新品种创新项目（PZCZ201718）作者简介：孙博文（1997-），男，河南许昌人，硕士，研究方向为蔬菜遗传育种与生物技术。（E-mail）2372765736@qq.com。黄耀为共同第一作者。通讯作者：钱春桃，(E-mail)chuntaoq@njau.edu.cn
更新日期/Last Update: 2022-07-07