籽用大麻性別連鎖標記的驗證及SCAR標記開發

趙銘森 方書生 陳瑤 康紅梅 高金虎 陳思遠 楊昕 馮旭平 張立武

摘 ?要??大麻是雌雄異株的異花授粉作物，在苗期鑒定出植株的性別，對大麻育種及生產具有重要的意義。為開發與籽用大麻品種性別連鎖的SCAR標記，本研究以汾麻3號、榆社麻和蒼山麻為試驗材料，研究已有的11個性別連鎖標記在籽用大麻品種中的適用性。結果表明，3個標記（OPD-05、UBC-354和MADC2）在籽用大麻品種中呈雄性特異，尚未發現雌性特異條帶。回收雄性特異條帶并測序，根據測序結果新開發了6個SCAR標記，其中MADC2-8表現最好;進而利用123份已知性別的樣本來檢測MADC2-8的準確性，發現其平均準確性高達98.34%。該研究不僅豐富了大麻性別鑒定的分子標記，而且為大麻性別研究和實際應用奠定基礎。

關鍵詞 ?大麻;籽用品種;性染色體;SCAR;MADC中圖分類號??S563.3??????文獻標識碼??A

Evaluation of Markers Linked to Sex-specific and Development of SCAR Makers in Seed Hemp （Cannabis sativa?L.）

ZHAO Mingsen^1，2， FANG Shusheng^2，3*， CHEN Yao^2，3， KANG Hongmei¹， GAO Jinhu¹， CHEN Siyuan^2，3， YANG Xin^2，3， FENG Xuping¹， ZHANG Liwu^2，3**

1.?Institute of Economic?Crops， Shanxi Academy of Agricultural Sciences， Taiyuan， Shanxi?030000， China; 2. Key Laboratory for Genetics， Breeding and Multiple Utilization of Crops，?Ministry of Education / Fujian Key Laboratory for Crop Breeding by Design， Fujian Agriculture and Forestry University， Fuzhou， Fujian?350002， China; 3.?Experiment Station of Jute and Kenaf in Southeast China， Ministry of Agriculture and Rural Affairs / Public Platform for Germplasm Resources of Bast Fiber Crops of Fujian， Fujian Agriculture and Forestry University， Fuzhou， Fujian?350002， China

Abstract ?Hemp?（Cannabis sativa L.） is a dioecious and cross pollinated plant. Sex identification of plants at seedling stage is of great significance for breeding and production in hemp. To develop new SCAR markers linked to sex-specific， three Chinese local seed hemp varieties， Fenma No.3， Yushema and Cangshanma， were used as the experimental materials to investigate the application of 11 markers linked to sex-specific in Chinese local seed hemp varieties in this study. The results showed that three makers （OPD-05 and UBC-354 as well as MADC2） were male-specific， while no female specific bands were found. Novel SCAR markers were developed on the basis of sequences of male specific bands. Among them， MADC2-8 was the best. Furthermore， 123 samples with known sex identification were used to assess the accuracy of MADC2-8. The average accuracy was 98.34%. These results?would?not only enrich the molecular markers linked to sex-specific， but also lay a foundation for the research and practical application of sex identification in hemp.

Keywords ?Cannabis sativa; seed variety; sex chromosome; SCAR; MADC

DOI10.3969/j.issn.1000-2561.2019.10.023

大麻（Cannabis sativaL.）為大麻科（Cannabinaceae）大麻屬（CannabisL.），屬短日照異花授粉作物，雌雄異株^[1]。按栽培目的進行分類，大麻可分為纖用型（工業大麻）、籽用型（油用型）、藥用型等。籽用型品種籽產量高，含油可達32%以上;纖用型品種麻皮產量高，出麻率高，纖維柔軟、色白、有光澤;藥用型品種花葉產量高，大麻二酚含量高^[1-4]。大麻植株有雌株及雄株不同類型，不同性別的植株形態存在較大差異。大麻從種子萌發到開花一般需要3個月以上的時間，在此之前不能從形態上準確判斷植株性別^[5]，只有在開花時才能從形態上準確判斷植株性別，生產上通常采用多留苗，到時雄性植株開花后自然死亡，這樣造成極大的浪費。……