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作物学报 ›› 2019, Vol. 45 ›› Issue (3): 335-343.doi: 10.3724/SP.J.1006.2019.82035

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

通过分子标记辅助选择将耐储藏主效QTL qSS-9 Kas转入宁粳4号提高其种子贮藏能力

张平1,姜一梅1,曹鹏辉1,张福鳞1,伍洪铭1,蔡梦颖1,刘世家1,田云录1,江玲1,*(),万建民1,2   

  1. 1南京农业大学作物遗传与种质创新国家重点实验室 / 农业部长江中下游粳稻生物学与遗传育种重点实验室 / 江苏省植物基因工程技术研究中心, 江苏南京 210095
    2中国农业科学院作物科学研究所 / 农作物基因资源与基因改良国家重大科学工程, 北京100081
  • 收稿日期:2018-07-06 接受日期:2018-12-24 出版日期:2019-03-12 网络出版日期:2019-01-03
  • 通讯作者: 江玲
  • 基金资助:
    本研究由国家转基因生物新品种培育重大专项(2016ZX08001006);江苏省重点研发项目(BE2018388);江苏省重点研发项目(BE2017368);江苏省现代作物生产协同创新中心项目资助

Introducing qSS-9 Kas into Ningjing 4 by molecular marker-assisted selection to improve its seed storage ability

Ping ZHANG1,Yi-Mei JIANG1,Peng-Hui CAO1,Fu-Lin ZHANG1,Hong-Ming WU1,Meng-Ying CAI1,Shi-Jia LIU1,Yun-Lu TIAN1,Ling JIANG1,*(),Jian-Min WAN1,2   

  1. 1 State Key Laboratory of Crop Genetics and Germplasm Enhancement / Key Laboratory of Biology, Genetics and Breeding of Japonica Rice in Mid-lower Yangtze River, Ministry of Agriculture / Research Center of Jiangsu Plant Gene Engineering, Nanjing Agricultural University, Nanjing 210095, Jiangsu, China
    2 National Key Facility for Crop Gene Resources and Genetic Improvement / Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2018-07-06 Accepted:2018-12-24 Published:2019-03-12 Published online:2019-01-03
  • Contact: Ling JIANG
  • Supported by:
    This study was supported by the National Major Project for Developing New GM Crops(2016ZX08001006);the Key Science and Technology Project of Jiangsu Province(BE2018388);the Key Science and Technology Project of Jiangsu Province(BE2017368);Jiangsu Collaborative Innovation Center for Modern Crop Production

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摘要：

利用置换系SL36作为qSS-9位点上Kasalath等位基因的供体亲本, 各方面农艺性状较好的宁粳4号作为轮回亲本, 通过回交和自交, 并使用4个与qSS-9紧密连锁的分子标记Y-10、Y-11、Y-14、Y-13进行基因型的检测筛选, 对宁粳4号进行耐贮性的遗传改良, 获得了既耐贮藏、大多农艺性状又接近于宁粳4号的较为稳定的优良新品系, 克服了宁粳4号耐贮藏性差的弱点。获得的新品系种子在人工老化和自然老化条件下均表现出发芽率明显提高、丙二醛含量显著降低、TTC染色效果更明显, 表明转入耐储藏主效QTL Qss-9 Kas的宁粳4号新品系耐贮性显著提高。

关键词: 水稻, 置换系, 分子标记辅助选择, 耐贮性, 遗传改良

Abstract:

In this paper, the chromosome segment substitution line SL36 was used as the donor parent with the Kasalath allele at the qSS-9 locus. Ningjing 4, a commercial cultivar with ideal agronomic traits, was used as a recurrent parent, through continuous self-pollination and backcrossing for four consecutive years. Four molecular markers Y-10, Y-11, Y-14, Y-13 closely linked to qSS-9 were used to screen genotypes, and molecular marker-assisted selection (MAS) was used for seed storage ability breeding for Ningjing 4. Through genetic improvement, we obtained inheritable lines with high seed storage ability. Most agronomic traits of the lines were nearly the same as those of Ningjing 4. These lines showed significantly higher germination rate, lower malondialdehyde content and more obvious TTC staining effects under artificial aging and natural aging conditions compared with Ningjing 4, indicating that the new lines with qSS-9 Kas have high seed storage ability.

Key words: rice, chromosome segment substitution line, molecular marker-assisted selection, seed storage ability, genetic improvement

图1

染色体片段置换系SL36的基因型示意图(A)和回交育种流程图(B)"

表1

本文中qSS-9的筛选标记"

标记
Marker		 正向引物
Forward primer (5°-3°)		 反向引物
Reverse primer (5°-3°)
Y-10 ACCTCAAGTTTTCCTATTAGT TAGAGTGACCTGCTAATGAG
Y-11 ACGATTAGTTCAGTCCTTACAC AACGGCTCAACGATCAGTAC
Y-14 AAAAAGGATGGGAAACTGACC TTTGAACTCAGTACCTTGGGG
Y-13 AAAAAGGATGGGAAACTGACC TACAAATAATCCCGATGCCG

图2

亲本及F1种子在自然环境下贮藏18个月后的种子发芽率和TTC法测定的种子生活力 A: 亲本及F1种子发芽率测定; B、C: 亲本及F1发芽表现和TTC法种子生活力表现, 从左到右依次是SL36、宁粳4号和F1。**表示与宁粳4号的发芽率相比, 在0.01水平上存在极显著差异。"

图3

各亲本和BC5F2在与qSS-9紧密连锁的分子标记Y-10、Y-11、Y-14、Y-13下的基因型鉴定 A: 利用与qSS-9紧密连锁的分子标记Y-10、Y-11、Y-14和Y-13对各亲本的基因型进行鉴定; 其中, 1: DNA marker; 泳道2~5、6~9、10~13和14~17分别代表各品系在Y-10、Y-11、Y-14和Y-13的基因型; 泳道2、6、10和14为Nipponbare; 泳道3、7、11和15为宁粳4号; 泳道4、8、12和16为置换系SL36; 泳道5、9、13和17为Kasalath。B: 利用与qSS-9紧密连锁的分子标记Y-10、Y-11、Y-14和Y-13对宁粳4号、SL36及BC2F1间的基因型进行鉴定: 其中1: DNA marker; 泳道2~6分别代表宁粳4号、SL36及BC2F1在Y-10下的基因型; 7~11分别代表宁粳4号、SL36及BC5F2在Y-11下的基因型; 12~16分别代表宁粳4号、SL36及BC2F1在Y-14下的基因型; 17~21分别代表宁粳4号、SL36及BC2F1在Y-13下的基因型。"

图4

亲本和后代种子在不同老化处理下的发芽率和丙二醛的含量 A: 宁粳4号、SL36、BC3F2 (Y6647)在自然老化一年MDA含量; B: 宁粳4号、SL36、BC3F2在自然老化一年和两年的发芽率; C: 亲本和BC5F1种子在人工老化处理35 d后的发芽率; D: 宁粳4号、Y3059-1-7、Y3059-1-8、Y3059-1-9的发芽率; E~H: 老化处理30 d后的发芽情况。E、F、G、H分别代表宁粳4号、Y3059-1-7、Y3059-1-8、Y3059-1-9。*表示在P = 0.05水平上差异显著; **表示在P = 0.01水平上差异极显著。"

图5

宁粳4号和Y3059的TTC试验、直链淀粉含量和株型的比较 A: 老化处理30 d后宁粳4号和Y3059的TTC染色情况, 从左到右分别代表宁粳4号、Y3059-1-7、Y3059-1-8、Y3059-1-9; B: Y3059及宁粳4号种子中TTCH的含量; C: 宁粳4号和Y3059老化处理前后表观淀粉含量的变化; D: 抽穗时期宁粳4号(左)和BC5F2 (右)株型; E: 抽穗35 d后宁粳4号(左)和BC5F2 (右)株型。*表示在P = 0.05水平上差异显著; **表示在P = 0.01水平上差异极显著。"

表2

宁粳4号和Y3059农艺性状的比较"

抽穗期
HD (d)		 株高
PH (cm)		 分蘖数
NT		 结实率
SSR (%)		 千粒重
TGW (g)		 单株产量
YPP (g)
宁粳4号Ningjing 4 87 ± 3 95 ± 4 11 ± 2 80.74 ± 0.02 25.50 ± 0.20 23 ± 0.18
BC5F2 (Y3059) 85 ± 3 107.5 ± 2.2** 13 ± 3* 83.08 ± 0.06 25.65 ± 0.90 22 ± 0.20
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