中國農業科學 ?? 2019, Vol. 52 ?? Issue (13): 2208-2219.doi: 10.3864/j.issn.0578-1752.2019.13.002

? 作物遺傳育種·種質資源·分子遺傳學 ? 上一篇    下一篇

利用BSA法發掘野生大豆種子硬實性相關QTL

陳靜靜,劉謝香,于莉莉,盧一鵬,張嗣天,張昊辰,關榮霞(),邱麗娟   

  1. 中國農業科學院作物科學研究所/國家農作物基因資源與遺傳改良重大科學工程/農業部種質資源利用重點實驗室,北京 100081
  • 收稿日期:2019-02-28 接受日期:2019-04-14 出版日期:2019-07-01 發布日期:2019-07-11
  • 通訊作者: 關榮霞 E-mail:[email protected]
  • 作者簡介:陳靜靜,E-mail:[email protected]
  • 基金資助:
    國家自然科學基金(31830066);中國農業科學院基本科研業務費(S2018QY03)

QTL Mapping of Hard Seededness in Wild Soybean Using BSA Method

CHEN JingJing,LIU XieXiang,YU LiLi,LU YiPeng,ZHANG SiTian,ZHANG HaoChen,GUAN RongXia(),QIU LiJuan   

  1. Institute of Crop Sciences, Chinese Academy of Agricultural Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement (NFCRI)/Key Laboratory of Germplasm Utilization, Ministry of Agriculture, Beijing 100081
  • Received:2019-02-28 Accepted:2019-04-14 Online:2019-07-01 Published:2019-07-11
  • Contact: RongXia GUAN E-mail:[email protected]

摘要:

【目的】野生大豆的硬實性是大豆遺傳改良利用中的重要限制因素。利用BSA法發掘與大豆種子硬實性相關的QTL,為野生大豆在大豆遺傳改良中的合理利用奠定基礎。【方法】利用栽培大豆中黃39與野生大豆NY27-38雜交構建F2和F7分離群體,從每個單株選取整齊一致的種子,取30粒種子置于鋪有一層濾紙的培養皿中,加入30 mL蒸餾水,25℃培養箱中暗處理4 h,設3次重復,分別統計每個培養皿中正常吸脹和硬實種子數。在F2群體中,選取22個正常吸脹單株(吸脹率>90%)和16個硬實單株(吸脹率<10%);在F7群體中,選取20個完全吸脹單株(吸脹率=100%)和20個完全硬實單株(吸脹率=0%),單株DNA等量混合,分別構建2個吸脹和2個硬實DNA池。利用259對在親本間有多態性的SSR標記對吸脹和硬實DNA池進行檢測,篩選在吸脹和硬實DNA池間表現多態性的SSR標記;用192個SSR標記檢測F7分離群體,構建遺傳圖譜,利用復合區間作圖法定位大豆硬實相關QTL。【結果】利用F2個體構建的吸脹和硬實DNA池,在第2染色體16.3 Mb區間和第6染色體23.4 Mb區間分別檢測到10個和8個在兩池間有差異的SSR標記。利用這些標記檢測F2群體,將第2染色體的QTL定位于Satt274與Sat_198間的276.0 kb區間,該區間包括已克隆的大豆硬實基因GmHs1-1,解釋17.2%的表型變異。第6染色體的QTL位于標記BARCSOYSSR_06_0993與BARCSOYSSR_06_1068間,可解釋17.8%的表型變異。利用F7株系構建的吸脹和硬實DNA池,在第2(27.4 Mb區間)、6(27.8 Mb 區間)和3染色體(18.2 Mb區間)分別檢測到11個、9個和4個在兩池間有多態性的SSR標記。利用F7群體構建包括192個SSR標記、覆蓋2 390.2 cM的遺傳圖譜,共檢測到3個硬實相關QTL,其中第2染色體定位到的QTL位于標記Satt274與Sat_198間,可解釋23.3%的遺傳變異。第6染色體定位到的QTL位于標記Sat_402與Satt557之間,可解釋20.4%的表型變異。在第3染色體標記Sat_266與Sat_236間發現一個可以解釋4.9%表型變異的QTL,與BSA法檢測的結果相符。【結論】利用BSA法可以檢測到傳統遺傳作圖定位的所有與硬實性相關的QTL,證明BSA法發掘大豆種子硬實性主要QTL的高效性。

關鍵詞: 大豆, 種子硬實, QTL定位

Abstract:

【Objective】 Hard seededness of wild soybean is an important effector that limits the utilization of wild resources in soybean genetic improvement. Bulked segregant analysis (BSA) was employed to identify major quantitative trait loci (QTLs) related with hard seededness in soybean, which laid a foundation for effective utilization of wild soybean germplasm in cultivated soybean improvement. 【Method】 F2 and F7 segregation populations were constructed from a cross between cultivated soybean Zhonghuang39 and wild soybean NY27-38. Uniformly sized seeds were selected from each line, and 30 seeds were soaked in a petri dish with 30 mL distilled water for 4 hours at 25℃. The assay was replicated 3 times. The number of permeable and impermeable seeds were counted. In F2 population, the first DNA pool was constructed from 22 individuals with permeable seeds (imbibition rate >90%), and second DNA pool was constructed from 16 individuals with impermeable seeds (imbibition rate <10%). In F7 population, 20 lines with permeable seeds (100% imbibition) and 20 lines with impermeable seeds (no imbibition) were used to construct two DNA pools, respectively. To detect genomic regions associated with hard seededness, these DNA bulks were genotyped with 259 polymorphic SSR markers to identify markers linked to QTL. A linkage map was constructed with 192 SSR markers, QTLs related with hard seededness were identified by composite interval mapping in F7 segregation population. 【Result】 Out of 259 SSR loci polymorphic between Zhonghuang39 and NY27-38, 10 and eight polymorphic SSR markers between the permeable and impermeable pools were detected in 16.3 Mb interval on chromosome 2 and 23.4 Mb interval on chromosome 6, respectively, in F2 population. The QTL region (276.0 kb) located between Satt274 and Sat_198 on chromosome 2 contained previously cloned gene GmHs1-1, the QTL explained 17.2% of the total genetic variation. The other QTL was mapped on chromosome 6 flanked by BARCSOYSSR_06_0993 and BARCSOYSSR_06_1068, accounting for 17.8% of the total genetic variation. In F7 population, eleven, nine and four SSR polymorphic markers between the permeable and impermeable pools were detected in 27.4 Mb interval on chromosome 2, 27.8 Mb interval on chromosome 6, 18.2 Mb interval on chromosome 3, respectively. A linkage map of 192 SSR markers and covering 2 390.2 cM was constructed through composite interval mapping in F7 population. Three QTLs related with hard seededness were detected. The QTL on chromosome 2 located between Satt274 and Sat_198, explained 23.3% of the total genetic variation; the QTL on chromosome 6 flanked by Sat_402 and Satt557, explained 20.4% of the total genetic variation; the QTL on chromosome 3 flanked by Sat_266 and Sat_236 accounted for 4.9% of the total genetic variation. 【Conclusion】 In this study, three QTLs related to soybean hard seededness were identified by both BSA and traditional linkage mapping, indicating that BSA is an effective strategy for identifying QTLs in soybean.

Key words: soybean, hard seededness, QTL mapping

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