發(fā)布時間：2018-04-29 15:37

  本文選題：白蠟 + 核心種質(zhì)��； 參考：《甘肅農(nóng)業(yè)大學(xué)》2014年碩士論文

【摘要】：白蠟是良好的鹽堿地造林樹種，用材樹種以及園林綠化樹種，構(gòu)建白蠟指紋圖譜對我國白蠟資源的充分合理利用、品種鑒定和知識產(chǎn)權(quán)保護均具有重要意義。本實驗利用SSR分子標記技術(shù)對白蠟種質(zhì)以及絨毛白蠟無性系進行了分析，篩選出了一批多態(tài)性豐富的SSR引物，構(gòu)建了SSR指紋圖譜并對白蠟種質(zhì)及絨毛白蠟無性系進行了遺傳多樣性評價。主要結(jié)果如下： 1．本研究建立了適合白蠟SSR-PCR反應(yīng)的最佳體系，用于白蠟SSR標記的進一步研究。本研究采用L16(45)正交設(shè)計和單因素試驗對影響白蠟SSR-PCR的Taq聚合酶用量、Mg2+濃度、DNA模板濃度、dNTP濃度和引物濃度等5個因素在4水平上進行篩選。優(yōu)化后的白蠟SSR反應(yīng)體系為：Mg2+(25mmol/L)0.8μL、引物(10μmol/L)0.2μL、Dntp (10mmol/L)0.3μL、Taq酶(5U/μL)0.05μL、DNA模板(10ng/μL)2.00μL、10×PCR緩沖液1.0μL，ddH2O5.45μL，總體積10.0μL。該結(jié)果為今后利用SSR-PCR標記技術(shù)研究分析白蠟奠定了基礎(chǔ)。 2．選用的12份白蠟核心種質(zhì)材料，，從156對SSR引物中篩選出10對反應(yīng)穩(wěn)定，擴增條帶清晰，多態(tài)性強的引物。10對引物共擴增出35個多態(tài)位點，且均為多態(tài)位點，多態(tài)性比例為100%。平均每對引物檢測到3.5個多態(tài)位點。多態(tài)性信息含量(PIC)變幅為0.3648～0.6725，平均為0.5480，引物20的PIC值最高，引物30的PIC值最低。利用引物42、18、26、30構(gòu)建引物組合，可將12個白蠟樹種完全區(qū)分開，每一份白蠟樹種都有自己獨特的指紋圖譜。UPGMA聚類及遺傳多樣性分析基本能將不同屬白蠟樹種分開，并能體現(xiàn)白蠟不同種間的親緣關(guān)系，結(jié)合白蠟SSR指紋圖譜，在白蠟種間鑒別、種質(zhì)資源管理、雜交育種和知識產(chǎn)權(quán)保護等方面具有重要的參考意義。 3．以46份絨毛白蠟無性系優(yōu)樹為實驗材料，在156對SSR引物中篩選出17對反應(yīng)穩(wěn)定，擴增條帶清晰，多態(tài)性強的引物。這17對引物共擴增出68個多態(tài)位點，且均為多態(tài)位點，多態(tài)性比例為100%。平均每對引物檢測到4.0個多態(tài)位點。PIC變幅為0.7059～0.1970，平均0.5184，引物42的PIC值最高，引物16的PIC值最低。利用引物12、16、E3、17、24、5、7、F3、4、42、C3、37、30構(gòu)建引物組合，可將46個絨毛白蠟無性系樹種完全區(qū)分開，每一份絨毛白蠟無性系樹種都有自己獨特的指紋圖譜。通過UPGMA聚類分析，不同性狀和株型的絨毛白蠟無性系大致能被區(qū)分開來，遺傳多樣性分析數(shù)據(jù)表明絨毛白蠟無性系遺傳多樣性比較豐富，說明本研究建立的評價體系具有參考意義。
[Abstract]:White wax is a good afforestation tree species in saline-alkali land. It is of great significance to construct the white wax fingerprint for the rational utilization of Chinese white wax resources, variety identification and intellectual property protection. In this study, SSR markers were used to analyze the white wax germplasm and fluffy white wax clones, and a number of polymorphic SSR primers were screened out. SSR fingerprint was constructed and genetic diversity of white wax germplasm and fluffy white wax clone was evaluated. The main results are as follows: 1. In this study, the optimum system for SSR-PCR reaction of white wax was established, which was used for further study of SSR labeling of white wax. In this study, the orthogonal design and single factor test were used to screen five factors, such as the concentration of Taq polymerase and the concentration of Taq template and primer, which affect the dosage of Taq polymerase and the concentration of primer. The optimized SSR reaction system was as follows: 10 渭 mol/L)0.2 / L Dntp 10 渭 mol / L 10 渭 mol / L Taq 5 U / 渭 L 0. 05 渭 L SSR template, 10 渭 L 10 渭 L 10 脳 PCR buffer 1. 0 渭 L LddH 2O 5.45 渭 L, and 10 渭 L 路L ~ (10) 渭 L ~ (10) 渭 L ~ (10) 渭 L ~ (10) 渭 L 路L ~ (-1) for 10 渭 L 路L ~ (-1), 10 渭 L ~ (10) 渭 L 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1). The results laid a foundation for the study and analysis of white wax by SSR-PCR labeling technique in the future. 2. Ten pairs of primers with stable reaction, clear amplified bands and strong polymorphic bands were selected from the 12 white wax core germplasm materials, and 35 polymorphic loci were amplified by 10 pairs of primers, all of which were polymorphic, and the proportion of polymorphism was 100%. An average of 3.5 polymorphic loci per pair of primers were detected. The variation of polymorphic information content (Pi) was 0.3648 ~ 0.6725 with an average of 0.5480. The PIC value of primer 20 was the highest, and the PIC value of primer 30 was the lowest. The primer combination was constructed by using primer 42O18 / 2630, and 12 species of white wax tree can be completely distinguished. Each species has its own unique fingerprint map. UPGMA clustering and genetic diversity analysis can basically separate the species of different genus of white wax tree. It can reflect the relationship between different species of white wax, combined with the SSR fingerprint of white wax, it has important reference significance in identification of white wax species, management of germplasm resources, cross breeding and intellectual property protection, etc. 3. Using 46 fluffy white wax clones as experimental materials, 17 pairs of primers with stable reaction, clear amplified bands and strong polymorphism were screened out of 156 pairs of SSR primers. A total of 68 polymorphic loci were amplified from 17 pairs of primers, all of which were polymorphic, with a polymorphic ratio of 100. The average number of polymorphic loci per primer was 0.70590.197, with an average of 0.5184.The PIC value of primer 42 was the highest, and the PIC value of primer 16 was the lowest. The primer combination was constructed by using the primer 12A16 E3C3C3C3730. The primer combination can completely distinguish 46 species of fluffy white wax asexual species, and each species has its own unique fingerprint map. The primer combination can be used to construct a primer combination of primer 12C16E3C3C3C3H730, and each of them has its own unique fingerprint map. The primer can be used to construct a primer combination. By UPGMA cluster analysis, different traits and plant types of fluff white wax clones can be roughly distinguished, and genetic diversity analysis data show that the genetic diversity of fluffy white wax clones is relatively rich. It shows that the evaluation system established in this study has reference significance.
【學(xué)位授予單位】：甘肅農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：S792.41

【參考文獻】

相關(guān)期刊論文 前10條

1 林道哲;董順文;劉朝輝;王軍;邱有榮;黃纓;李首成;;26份甘藍型油菜自交系指紋圖譜分析[J];安徽農(nóng)業(yè)科學(xué);2008年20期

2 萬平,劉大鈞;SSR_S標記與植物遺傳育種研究(綜述)[J];安徽農(nóng)業(yè)大學(xué)學(xué)報;1998年01期

3 李擁軍,蘇加楷;苜蓿地方品種遺傳多樣性的研究──RAPD標記[J];草地學(xué)報;1998年02期

4 海林,王克晶,楊凱;半野生大豆種質(zhì)資源SSR位點遺傳多樣性分析[J];西北植物學(xué)報;2002年04期

5 李艷;;白蠟樹屬樹種的園林應(yīng)用探討[J];湖北林業(yè)科技;2006年01期

6 宋順華;鄭曉鷹;;甘藍品種的AFLP指紋鑒別圖譜分析[J];分子植物育種;2006年S1期

7 陳方永;倪海枝;何理坤;王引;盧國耀;王冬米;任正初;;浙江部分柿屬植物鑒定及親緣關(guān)系的SSR分析[J];果樹學(xué)報;2011年01期

8 郭小平,趙元明,劉毓俠;SSR技術(shù)及其在植物遺傳育種中的應(yīng)用[J];華北農(nóng)學(xué)報;1998年03期

9 吳莉英;唐前瑞;李達;陳麗;賀蘇華;王慧穎;尹恒;;非洲菊的AFLP指紋圖譜構(gòu)建[J];湖南林業(yè)科技;2008年04期

10 郭爾慶,祝鳳寶,王培忠;水曲柳無性系種子園營建技術(shù)的研究[J];吉林林業(yè)科技;2001年06期

本文編號：1820543