【作者】 周红伟；

【导师】 杜德志；

【作者基本信息】 青海大学 ， 作物遗传与育种， 2010， 硕士

【摘要】 我省低海拔地区主要种植甘蓝型油菜,品种多达7个,其中包括杂交种6个杂交种和常规种1个常规种。品种真伪及杂交种种子纯度鉴定,对于打击假冒伪劣,维护育种者的权益和保护农民的利益具有重要的意义。品种纯度的鉴定,传统的方法是采用田间种植鉴定,该方法可提供的检测指标有限,且准确性差。近年来发展的分子标记技术,能直接反映基因组水平的差异,已广泛应用到农作物品种指纹图谱构建和品种鉴定中,并成为杂交种真伪鉴定的有效手段。本研究应用SSR分子标记对我省目前广泛推广的甘蓝型油菜品种的指纹图谱及青杂系列杂交种种子纯度的快速鉴定技术进行了研究,得到以下结果:(1)从140对SSR引物中筛选出13对多态性较好的引物用于指纹图谱的构建,发现同时采用P013、P052这2对引物组合或同时采用P086、P027和P021这3对引物组合,可以将14份供试材料(包括6个杂交种品种、7个亲本材料及1个常规种)完全区分开来。这些指纹可用于亲本材料和杂交种真伪的鉴别。(2)筛选出在各品种中具有共显性标记的引物:青杂2号(P019、P027),青杂3号(P013、P027、P052),青杂4号(P027、P052、P086),青杂5号(P030、P085)。(3)利用获得的共显性引物对2008年大田生产杂交种的纯度进行鉴定,同时也采用传统田间种植的方法对杂交种进行纯度鉴定。实验室检测各杂交种纯度分别为:青杂2号纯度:83.41%(P027)、83.90%(P019);青杂3号纯度:87.02%(P027)、86.54%(P052)、86.06%(P013);青杂4号纯度:70.67%(P027)、69.23%(P052)、68.75%(P086);青杂5号纯度:85.37%(P030)、82.93%(P085)。田间鉴定各杂交种纯度分别为:青杂2号88.29%、青杂3号91.35%、青杂4号72.12%、青杂5号87.80%。实验室分子标记检测结果与田间种植鉴定结果趋势基本一致。但实验室分子标记检测结果均较田间鉴定结果偏低,造成这种差异的原因主要是田间鉴定以育性为依据,即田间表现可育为真杂种、田间表现不育为假杂种,而分子标记检测不仅可以区分可育与不育单株,而且还可以将可育株中的真杂种和恢复系单株、保持系单株及杂单株区分开。(4)将田间表现为可育而分子标记检测为父本带型(推测为恢复系)、田间表现为可育而分子标记检测为母本带型(推测为保持系)的单株分别与不育系测交,并自交,在云南鉴定测交F1和自交种的育性,结果表明推测为保持系单株的测交F1全为不育株,自交种全部可育;而推测为恢复系单株的测交F1全为可育株,自交种全部可育。证明了分子标记鉴定结果的准确性,同时也说明,相对于传统的田间种植鉴定方法,采用SSR分子标记进行杂交种纯度的鉴定更为准确。更多还原

【Abstract】 In lower altitude areas of Qinghai Province where mainly planting Brassica napus, there are seven varieties that contains six hybrids, one conventional variety. identificate Hybrids authenticity and purity rapidly that it is important for fight against counterfeiting and protected the interests of breeders and farmers has grateful significance. In the past mainly testing purity use morphological in the field and that can offer limited detection targets, morphological identification way can not adapt for purity analysis more and more. Molecular markers have been developed in recent years, this way reflection the differences of genomic directly that has been applied to constructed agriculture crops and varieties fingerprinting and identification widely, that is an effective mean for hybrids authenticity identification. In this study, application of SSR molecular markers constructed 14 Cultivars spring Brassica napus fingerprints of Qinghai and study on Qingza lines hybrids rapid purity identification way, as the following results:(1) 13 pairs polymorphic primers screened from 140 pairs SSR primers that using Construction fingerprints showed that the primer combinations P013 + P052 or P086 + P027 + P021 amplification bands can be completely distinguished14 materials that contains 4 hybrids and its parents, Huaxie No1 hybrids,Hufeng010 hybrids and Qingyou No14. Those fingerprints can be used for parent materials and hybrids authenticity identification.(2) Selected SSR primers have co-dominant bands that adapt to varieties purities identification. Qingza No.2 hybrid (P019、P027), QingzaNo.3 hybrid (P013、P027、P052),Qingza No. 4 hybrid(P027、P052、P086) ,Qingza No.5 hybrid (P030、P085).(3) Using above primers identified the hybrids purities that production of 2008, by field cultivation methods identified those hybrids Purity meanwhile. Laboratory testing hybrid purity: Qingza 2 Purity:83.41%(P027)、83.90%(P019);Qingza No. 3 purity:87.02%(P027)、86.54%(P052)、86.06%(P013);Qingza No. 4 purity:70.67%(P027)、69.23%(P052)、68.75%(P086);Qingza No. 5 purity:85.37%(P030)、82.93%(P085).Respectively, the field identification purities were 88.29%, 91.35%, 72.12%, 87.80%;through comparison lab. and field purities identification results, indicated that SSR markers tested purities all lower than field purities, that because of field planting identification purity according for saxes, and SSR marker not only distinct sterile from hybrids, but also districted more Restorer, and other rape plants in the field.(4) 2009 winter in Yunnan tested materials that from Qingza No. 2 field tested hybrids, lab. tested sterile bands and restorer bands plants that crossed with true sterile, and tested self crossed seeds that from those plants self cross. Indicated that those hybrids from lab. tested sterile bands plants crossed with true sterile were all true sterile, self cross seed were all maintainers, and hybrids from lab. tested restorer bands plants crossed with true sterile were all true hybrids, self cross seed were all restorer. Indicated that SSR marker testing hybrid purity was correct than field plant testing.更多还原