利用RAPD技术对吉林省重要果树资源的鉴定研究
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摘要
利用RAPD技术对小苹果、越橘、穗醋栗、李、杏、樱、猕猴桃、抗寒葡萄、梨等68份吉林省果树种质在资源分类、亲缘关系及种质鉴别等方面进行了研究。
     1、用10个引物对吉林省8份小苹果种质进行RAPD扩增,结果表明:用S337,S443,S446这3个引物可完全鉴别这8份小苹果种质资源。K9、大秋和龙冠聚为一类,新苹1号与金红聚为一类,黄太平、吉早红、和山定子各自聚为一类。金红与吉早红二个品种不仅在RAPD条带上表现了很大的差异,而且遗传距离为0.295455,证明吉早红与金红之间亲缘关系较远。
     2、用7个引物对吉林省7份越橘种质进行RAPD扩增,结果表明:越橘的RAPD扩增有较强的多态性,共扩增出42条带,多态性在33.3%~100%之间。用2个引物就可以很好地区别和鉴定供试的7个越橘品种。
     3、用12个引物对吉林省10份黑穗醋栗种质进行RAPD扩增,结果表明:黑穗醋栗RAPD条带在600bp~3200bp之间,DNA多态性较强,共扩增出58条带,多态性在50%~100%之间。用S84与S89可鉴定所有品种;用S84与S155也可鉴定10个品种。
     4、用12个引物对以吉林省11份李、杏、樱种质进行RAPD扩增,结果表明:共扩增出李、杏、樱80条带,多态性在66.7%~100%之间。由各引物扩增结果来看,李、杏、樱的扩增条带有明显差异,产生条带的多寡及区域不尽相同,有的多态性条带为杏、樱独有,而李少有或没有。
     5、选用10个随机引物对11份猕猴桃种质进行RAPD扩增,结果表明:每个引物扩增带在4~10个之间,DNA长度大小从400bp~3000bp。共扩增64条带,DNA多态性强,在60%~100%,平均为79.9%。S6-2700,S19-1450为软枣猕猴桃特有带,S3-2000, S21-2000为狗枣猕猴桃特有带, S21-S3-800,S45-700,S43-1200为葛枣猕猴桃特有带。
     6、、用9个随机引物对10份抗寒葡萄品种(品系)进行RAPD扩增,结果表明:山葡萄种质的DNA多态性很强,说明遗传变异很丰富,然而山葡萄种质间多态性不是很强。从各种质间相似系数来看,山葡萄群体间相似系数都在0.6以上,说明它们亲缘关系近。起源于俄罗斯的RS自聚为一组,说明其与东北山葡萄在地理起源上较远。
     7、用9个引物对11份梨种质进行RAPD扩增,结果表明:梨的RAPD扩增结果获得了较强的多态性,在25%~100%之间,有一定的DNA多态性,扩增的条带数为4~9条,平均5.18条,扩增片段大小在300~4800bp之间。S38-500,S89-1200,S91-1800,S91-900,S93-1200,S93-600,S81-600,S82-1000,S86-1500,S86-1000,S86-900,为各种质的共有带。利用引物S23、S37、S38可鉴别供试梨种质资源。
In this thesis, we studied the classification, genealogy and idioplasm distinction of 68 kinds of fruit trees using of RAPD technique.
     1. We analysised 8 samples of small apples using RAPD amplification with 10 pairs of specific primers. The result shows that the primers S337, S443 and S446 are competent to identify the eight breeds of small apples. K9、Daqiu and long guanju can be classified into same group , No.1 of Xingping and Jinhongju . Huangtaiping, Jinzaohong and Shandingzi bear their own catogories respectively, in this study, Jinhong and Jizaohong performe differently in RAPD bands, and have a genetic distance of 0.295455, which shows that Jizaohong and Jinhong have distant relative relationship.
     2. We analysised 7 samples of blueberries using RAPD amplification with 7 pairs of primers in Jilin province. The result shows great varieties in RAPD of blueberrie , as 42 bands are amplified, which means a variety between 33.3% and 100%. With 2 primers, we can distinguish and identify the blueberries varieties.
     3. We analysised 10 samples of small apples using RAPD amplification with 12 pairs of specific primers in Jilin province. The result shows that the size of RAPD bands of black currants are between 600bp and 3200bp. Its RAPD has greater polymorphism, and 58 bands are amplified, which mean a polymorphism between 50% and 100%. S84 and S89 can be used to identify the whole 10 samples. a S84 and S155.
     4. We analysised the eleven groups of plums, apricots and cherries to using RAPD with 12 pairs of primers in Jilin province. The result shows that 80 bands are produced in amplifying plums, apricots and cherries and its polymorphism is between 66.7% and 100%. According to the results with all the primers, the amplified stripes of plums, apricots and cherries have distinct differences and the amount and region for the amplified stripes are different. Some polymorphic stripes are exclusive to the apricots and cherries and the plums have less or no stripes.
     5. We choose 10 primers at random to amplify the RAPD of the kiwi fruits. It shows that the amplified stripes numbers of each primer are between 4 and 10 and the lengths of DNA range from 400bp to 3000bp, altogether 64 stripes are amplified. The DNA has polymorphism is between 60% and 100% , with an average of 79.9%. S6-2700 and S19-1450 are special stripes to soft date kiwi fruits. S3-2000 , S21-2000 are the special stripes to Gou dates kiwi fruits , S21-S3-800 , S45-700 and S43-1200are the special stripes for Ge dates kiwi fruits .
     6. We choose 9 pairs of primers at random to amplify the RAPD of ten varieties of hill grapes. It shows that the hill grapes’DNA has greater polymorphism, which means an abundance of the genetic variation, but the polymorphism among these varieties is not very big. Seeing from the similar coefficients among these varieties, the similar coefficients among these varieties is above 0.6, which means they have close relative relationships. The RS from Russia are in the same group, which means the geographical origin between RS and the northeast mountain grapes is remote.
     7. We choose 9 primers at random to amplify the RAPD of 11 groups of pears. The result of RAPD amplification of pear shows that it has gteater polymorphism which is between 25% and 100% and have certain DNA polymorphism. The amplified stripes are between 4 and 9 with an average of 5.18. The sizes of the amplified fragments are between 300bp and 4800bp. S38-500, S89-1200, S91-1800, S91-900, S93-1200, S93-600, S81-600, S82-1000, S86-1500, S86-1000 and S86-900 are the common stripes of different varieties. Primers S23, S37 and S38 can identify the qualities of the pear resources.
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