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园艺学报 ›› 2014, Vol. 41 ›› Issue (3): 409-416.

• 果树 •    下一篇

柑橘10 个品种实生后代多倍体的发掘及SSR 鉴定

梁武军,解凯东,郭大勇,谢宗周,徐  强,伊华林,郭文武*   

  1. 华中农业大学园艺植物生物学教育部重点实验室,农业部华中地区园艺作物生物学与种质创制重点实验室,武汉 430070
  • 出版日期:2014-03-25 发布日期:2014-03-25
  • 基金资助:

    国家自然科学基金项目(31125024);国家‘863’计划课题(2011AA100205);教育部创新团队项目(IRT13065);湖北省
    自然科学基金创新群体项目(2013CFA001)

Spontaneous Generation and SSR Characterization of Polyploids from Ten Citrus Cultivars

LIANG Wu-jun,XIE Kai-dong,GUO Da-yong,XIE Zong-zhou,XU Qiang,YI Hua-lin,and GUO Wen-wu*   

  1. Key Laboratory of Horticultural Plant Biology(Ministry of Education),Key Laboratory of Central China Horticultural
    Crop Biology and Germplasm Enhancement(Ministry of Agriculture),Huazhong Agricultural University,Wuhan 430070,
    China
  • Online:2014-03-25 Published:2014-03-25

摘要: 秋季采摘柑橘成熟果实并剥取种子,将小粒种籽和瘪籽播于MT 培养基,待萌发长出2 ~ 3
片真叶时,取其叶片检测倍性;将饱满种籽播种于露地,对其幼苗首先通过形态筛选获得似多倍体的植
株,然后取其叶片用流式细胞仪进行倍性检测;最后对获得的多倍体植株进行SSR 分子鉴定。试验结果
表明,获得早金甜橙、卡特夏橙、塔罗科血橙、日辉橘、Ortanique 橘橙、默科特橘橙、华农本地早橘、
朱红橘和Flame 葡萄柚等9 个品种的四倍体分别为6、1、3、12、3、6、2、1 和10 株;获得早金甜橙、
塔罗科血橙、Itabori 甜橙和Ortanique 橘橙等4 个品种的三倍体分别为1、1、2、4 株。用13 对多态性SSR
引物对胚抢救获得的三倍体和四倍体的来源进行鉴定,发现早金甜橙、Itabori 甜橙的三倍体再生植株为异
源三倍体;塔罗科血橙、Ortanique 橘橙的三倍体可能为同源三倍体;早金甜橙和Ortanique 橘橙的四倍体
可能为同源四倍体;塔罗科血橙四倍体为异源四倍体。采用37 对多态性引物对从饱满种籽中获得的8 个
品种四倍体的来源进行鉴定,发现均为同源四倍体。这些自然发掘的多倍体资源对柑橘无核育种和相关
基础研究具有重要价值。

关键词: 柑橘, 多倍体, SSR 分子标记

Abstract: Seeds were extracted from mature fruits and undeveloped seeds were cultured in vitro and
mature seeds were sowed in field. Seedlings with 2–3 true leaves after germination were analyzed by
flow cytometry to screen polyploids,and simple sequence repeat (SSR) markers were used to characterize
the genetic origin of these polyploids. The results showed that a total of 44 tetraploid seedlings were obtained from all studied diploid genotypes,i.e. 6,1,3,12,3,6,2,1 and 10 seedlings from Early Gold
sweet orange,Cutter Valencia,Tarocco blood orange,Sunburst mandarin,Ortanique tangor,Murcott
tangor,Huanong Bendizao tangerine,Zhuhongju tangerine and Flame grapefruit respectively. A total of 8
triploid seedlings were obtained from all studied diploid genotypes,i.e. 1,1,2,4 seedlings from Early
Gold sweet orange,Tarocco blood orange,Itabori sweet orange and Ortanique tangor respectively. The
genetic origin of polyploids obtaining by embryo rescue were determined by 13 SSR markers,and the
results showed that the regenerated triploid plants of Early Gold and Itabori sweet oranges were
allotriploid and that of Tarocco blood orange and Ortanique tangor maybe were autotriploid. The
regenerated tetraploid plants of Early Gold sweet orange and Ortanique tangor maybe were autotetraploid
and those of Tarocco blood orange were allotetraploid. SSR analysis validated that the band profiles of
tetraploids derived from other eight diploid cultivars were identical to their diploid maternal line. These
exploited natural polyploids hold great value for both citrus seedless breeding and basic research.

Key words: Citrus, polyploidy, SSR(Simple Sequence Repeat)

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