小果木瓜型南瓜抗矮南瓜黃化嵌紋病毒分子標誌之建立

Simulation Study of Genomic Developing the Molecular Markers for Zucchini yellow mosaic virus Resistance in Winter Squash (Cucurbita moschata)

黃昭欽(Chao-Chin Huang) ； 歐尚靈(Shang-Ling Ou) ； 陳思勻(Sz-Yun Chen) ； 林崇正(Chong-Zheng Lin) ； 周汝潔(Ju-Chieh Chou) ； 古新梅(Hsin-Mei Ku)

《作物、環境與生物資訊》 11卷1期 (2014/03) Pp. 26-37

https://doi.org/10.30061/CEB.201403_11(1).0003

摘要

南瓜（Cucurbita spp.）為葫蘆科作物，果肉及種子極具營養價值。矮南瓜黃化嵌紋病毒（Zucchini yellow mosaic virus；ZYMV）是目前臺灣葫蘆科作物的主要病毒病害之一，其病徵包括植株生長遲緩，葉片黃化、斑駁、嵌紋及果實畸形，生長初期若遭感染，會嚴重影響產量。本研究選擇同為Cucurbita moschata的抗病種原Nigerian Local（NL）與臺灣市面上常見的感病種原小果木瓜型南瓜（Local Papaya；PY）作為親本。將F2和BC1子代接種ZYMV-TN3病毒株後，觀察子代族群病徵的分離比，推測ZYMV之抗性可能由兩個顯性互補基因調控。為尋找南瓜中與ZYMV抗性基因座連鎖的分子標誌，利用逢機增幅多型性DNA（Randomly Amplified Polymorphic DNA；RAPD）配合群聚分離族群分析（bulk segregation analysis；BSA）進行篩選，共找到7個可增幅出多型性片段的引子，將其分別轉換成序列特徵增幅區域（Sequence Characterized Amplified Region；SCAR）或切割增幅多型性序列（Cleaved Amplified polymorphic sequence；CAPS）分子標誌，於F2子代進行分析後，顯示CA83、CA215及SC522945與ZYMV抗性基因座連鎖。未來期望本試驗所得到的分子標誌可提高篩選效率及縮短育成抗病品種的時程。

關鍵字

南瓜 ； 矮南瓜黃化嵌紋病毒 ； 抗性基因座 ； 群聚分離族群分析 ； 逢機增幅多型性DNA ； 序列特徵增幅區域 ； 切割增幅多型性序列

並列摘要

Squash (Cucurbita spp.) has been grown by humans for fruits and seeds over 10,000 years. There are several viruses infecting squashes in Taiwan, and Zucchini yellow mosaic virus (ZYMV) is one of the most destructive viruses. Breeding for ZYMV resistance in squash is the best way to control it. The purpose of this study is to tag molecular markers which show the linkage to ZYMV resistance. A ZYMV resistant line, ”Nigerian Local (NL)”, was crossed to susceptible line, ”Local Papaya (PY)”and F2, BC1 and BC4 populations were generated for genetic analysis and marker tagging. The result suggested that the inheritance of ZYMV resistance in the near isogenic lines derived from PY x (PY x NL) is controlled by two dominant complementary genes. A total of 631 random amplified polymorphic DNA (RAPD) and bulk segregant analysis (BSA) were performed to screen useful linked markers. Seven polymorphic RAPD fragments were converted into sequence characterized amplified region (SCAR) or cleaved amplified polymorphic sequence (CPAS) markers. After linkage analysis, only three markers were shown linkage to ZYMV resistance. In the future, these markers would be useful in marker-assisted selection (MAS) and help us to select the resistant plants more efficiently.

並列關鍵字

Cucurbita spp. ； ZYMV ； resistance locus ； BSA ； RAPD ； SCAR ； CAPS