桂野生山金柑的倍性分析和SSR分子鉴定
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摘要
【目的】研究‘桂野生山金柑’的遗传学特性,为柑橘种质资源的鉴定及创新利用提供理论依据。【方法】采用倍性分析和SSR标记相结合,对‘桂野生山金柑’进行遗传鉴定分析。【结果】倍性分析结果表明‘桂野生山金柑’是二倍体;从24对SSR引物中筛选出6对重复性好、条带清晰、多态性强的核基因组引物,在供试材料中获得等位基因条带共30条,其中具有多样性差异条带25条,多态性比例为79.17%。聚类分析的结果表明不同供试材料的遗传相似系数在0.600~0.802。【结论】‘桂野生山金柑’具有遗传特异性,不同于其他地区起源的山金柑或金柑,是一种新的柑橘种质资源。
【Objective】In order to provide a theoretical basis for the identification and utilization of Gui Wild Shanjingan,its genetic characteristics was studied【Methods】Using ploidy analysis and SSR molecular marker to analyze the genetic diversity of Gui Wild Shanjingan.【Results】The ploidy analysis result showed that Gui Wild Shanjingan was diploid. 6 pairs of SSR primers with good repeatability,high polymorphism and clear bands were screened from the 24 pairs of SSR primers. A total of 30 allele stripes were successfully observed,among them,25 bands were polymorphism. The polymorphic rate was 79. 17 %. Genetic similarities caculated by cluster analysis ranged from 0.600-0. 802. 【Conclusion】Gui Wild Shanjingan was significant different from Shanjingan or Jingan originated from other origions,it was a new cultivar with genetic specificity.
【Objective】In order to provide a theoretical basis for the identification and utilization of Gui Wild Shanjingan,its genetic characteristics was studied【Methods】Using ploidy analysis and SSR molecular marker to analyze the genetic diversity of Gui Wild Shanjingan.【Results】The ploidy analysis result showed that Gui Wild Shanjingan was diploid. 6 pairs of SSR primers with good repeatability,high polymorphism and clear bands were screened from the 24 pairs of SSR primers. A total of 30 allele stripes were successfully observed,among them,25 bands were polymorphism. The polymorphic rate was 79. 17 %. Genetic similarities caculated by cluster analysis ranged from 0.600-0. 802. 【Conclusion】Gui Wild Shanjingan was significant different from Shanjingan or Jingan originated from other origions,it was a new cultivar with genetic specificity.
引文
[1]邓秀新.中国柑橘品种[M].北京:中国农业出版社,2008:190.
[2]陈杰忠.果树栽培学各论[M].北京:中国农业出版社,2003:4.
[3]李怀福,胡小三.山金柑实生苗童期的研究[J].特产研究,2005(1):23-25,39.
[4]耿攀,阮小凤,杨勇,等.柿属植物种质资源遗传多样性的SSR分析[J].西北农林科技大学学报(自然科学版),2010,38(12):190-196.
[5]潘丽梅,朱建华,秦献泉,等.广西龙荔遗传多样性的ISSR分析[J].果树学报,2011,28(5):814-819.
[6]陈娇,王小蓉,杨浩茹,等.基于SSR标记的四川野生中国樱桃遗传多样性和居群遗传结构分析[J].园艺学报,2013,40(2):333-340.
[7]赵爽,苏淑钗,陈志钢,等.基于SSR的辽宁铁岭地区平榛遗传多样性与居群遗传结构分析[J].果树学报,2016,33(1):24-33.
[8]罗兵,孙海燕,徐港明,等.SSR分子标记研究进展[J].安徽农业科学,2013,41(12):5210-5212,5246.
[9]Ferrante S P,Lucretti S,Reale S,et al.Assessment of the origin of new citrus tetraploid hybrids(2n=4x)by means of SSR markers and PCR based dosage effects[J].Euphytica,2010,173(2):223-233.
[10]Amar M H,Biswas M K,Zhang Z W,et al.Exploitation of SSR,SRAP and CAPS-SNP markers for genetic diversity of Citrus germplasm collection[J].Scientia Horticulturae,2011,128(3):220-227.
[11]张连峰,何建,冯焱,等.金柑属及其近缘属植物亲缘关系的SSR分析[J].果树学报,2006,23(3):335-338.
[12]陈鹏.中国野生山金柑种质资源调查及遗传多样性研究[D].华中农业大学硕士学位论文,2011:17-27.
[13]邓秀新.国内外柑橘产业发展趋势与柑橘优势区域规划[J].广西园艺,2004(4):6-10.
[14]张俊娥,刘继红,邓秀新.采用倍性分析仪鉴定柑橘愈伤组织的遗传变异[J].遗传学报,2003,30(2):169-174.
[15]谢宗周,邓秀新,陈腾士,等.“桂橙一号”的遗传鉴定[J].南方园艺,2010,21(3):3-6.
[16]程运江,伊华林,庞晓明,等.几种木本果树DNA的有效提取[J].华中农业大学学报,2001,20(5):481-483.
[17]Bassam B J,Caetano-Anollés G,Gresshoff P M.Fast and sensitive silver staining of DNA in polyacrylamide gels[J].Analytical Biochemistry,1991,196:80-83.
[18]刘勇,吴波,刘德春,等.江西柑橘地方品种资源及野生近缘种SSR分子标记[J].江西农业大学学报,2005,27(4):486-490.
[19]刘通,邓崇岭,程玉芳,等.利用SSR和SRAP技术研究广西柑橘种质遗传多样性[J].华中农业大学学报,2016,35(2):23-29.
[20]廖振坤.湖南柑橘种质资源评价及主要病害分子鉴定[D].湖南农业大学博士学位论文,2007.
[21]沈丽娟,高克义,李志宏,等.发挥广西柑桔优势之浅见[J].广西园艺,2007(1):8-10.
[22]张建成.红肉脐橙类胡萝卜素合成酶基因(Cs PYS、Cs LCYB)功能分析与crt B转基因对柑橘类胡萝卜素合成的影响[D].华中农业大学博士学位论文,2009:71-72
[23]刘建军,陈克玲,胡强,等.特色地方柑橘资源“资阳香橙”的初步研究[J].西南农业学报,2008,21(6):1658-1660.
[24]刘冰浩,邓崇岭,陈传武,等.广西地方柑橘资源的ISSR分析[J].果树学报,2015,32(6):1001-1006.
[2]陈杰忠.果树栽培学各论[M].北京:中国农业出版社,2003:4.
[3]李怀福,胡小三.山金柑实生苗童期的研究[J].特产研究,2005(1):23-25,39.
[4]耿攀,阮小凤,杨勇,等.柿属植物种质资源遗传多样性的SSR分析[J].西北农林科技大学学报(自然科学版),2010,38(12):190-196.
[5]潘丽梅,朱建华,秦献泉,等.广西龙荔遗传多样性的ISSR分析[J].果树学报,2011,28(5):814-819.
[6]陈娇,王小蓉,杨浩茹,等.基于SSR标记的四川野生中国樱桃遗传多样性和居群遗传结构分析[J].园艺学报,2013,40(2):333-340.
[7]赵爽,苏淑钗,陈志钢,等.基于SSR的辽宁铁岭地区平榛遗传多样性与居群遗传结构分析[J].果树学报,2016,33(1):24-33.
[8]罗兵,孙海燕,徐港明,等.SSR分子标记研究进展[J].安徽农业科学,2013,41(12):5210-5212,5246.
[9]Ferrante S P,Lucretti S,Reale S,et al.Assessment of the origin of new citrus tetraploid hybrids(2n=4x)by means of SSR markers and PCR based dosage effects[J].Euphytica,2010,173(2):223-233.
[10]Amar M H,Biswas M K,Zhang Z W,et al.Exploitation of SSR,SRAP and CAPS-SNP markers for genetic diversity of Citrus germplasm collection[J].Scientia Horticulturae,2011,128(3):220-227.
[11]张连峰,何建,冯焱,等.金柑属及其近缘属植物亲缘关系的SSR分析[J].果树学报,2006,23(3):335-338.
[12]陈鹏.中国野生山金柑种质资源调查及遗传多样性研究[D].华中农业大学硕士学位论文,2011:17-27.
[13]邓秀新.国内外柑橘产业发展趋势与柑橘优势区域规划[J].广西园艺,2004(4):6-10.
[14]张俊娥,刘继红,邓秀新.采用倍性分析仪鉴定柑橘愈伤组织的遗传变异[J].遗传学报,2003,30(2):169-174.
[15]谢宗周,邓秀新,陈腾士,等.“桂橙一号”的遗传鉴定[J].南方园艺,2010,21(3):3-6.
[16]程运江,伊华林,庞晓明,等.几种木本果树DNA的有效提取[J].华中农业大学学报,2001,20(5):481-483.
[17]Bassam B J,Caetano-Anollés G,Gresshoff P M.Fast and sensitive silver staining of DNA in polyacrylamide gels[J].Analytical Biochemistry,1991,196:80-83.
[18]刘勇,吴波,刘德春,等.江西柑橘地方品种资源及野生近缘种SSR分子标记[J].江西农业大学学报,2005,27(4):486-490.
[19]刘通,邓崇岭,程玉芳,等.利用SSR和SRAP技术研究广西柑橘种质遗传多样性[J].华中农业大学学报,2016,35(2):23-29.
[20]廖振坤.湖南柑橘种质资源评价及主要病害分子鉴定[D].湖南农业大学博士学位论文,2007.
[21]沈丽娟,高克义,李志宏,等.发挥广西柑桔优势之浅见[J].广西园艺,2007(1):8-10.
[22]张建成.红肉脐橙类胡萝卜素合成酶基因(Cs PYS、Cs LCYB)功能分析与crt B转基因对柑橘类胡萝卜素合成的影响[D].华中农业大学博士学位论文,2009:71-72
[23]刘建军,陈克玲,胡强,等.特色地方柑橘资源“资阳香橙”的初步研究[J].西南农业学报,2008,21(6):1658-1660.
[24]刘冰浩,邓崇岭,陈传武,等.广西地方柑橘资源的ISSR分析[J].果树学报,2015,32(6):1001-1006.