摘要
为探寻甘蓝型油菜黄黑籽之间的种皮色泽表达差异的分子基础,探索cDNA-SRAP差异显示的可行性,选用培育7年的重组自交系群体(RILs),根据群体分离分析法(bulkedsegregantanalysis,BSA)构建了种子的RNA混合池,利用cDNA-SRAP法进行了差异显示研究。996对SRAP引物组合扩增出2100条带,2次扩增重复率为65.2%,其中在黄黑籽材料之间重复稳定出现的差异片段有12条,长度在100～300bp之间。选择其中7条差异片段,进行回收、克隆和测序,发现2个片段分别与拟南芥的NAD+ADP核糖转移酶及小肽转移蛋白3具有很高的同源性。结果表明,利用SRAP方法可以对甘蓝型油菜cDNA混合池进行差异显示研究,2条差异片段可能与甘蓝型油菜的种皮色泽基因表达相关。
The yellow-seed and black-seed in Brassica napus L.have different characters and different mechanisms in the seed color formation.The objective of this research was to explore the molecular mechanism of the differential expression between yellow and black immature seeds and demonstrate the feasibility of cDNA-SRAP for differential display.On the basis of the different lines with yellow and black seed-coat from recombinant inbred lines（RILs）which had self-pollinated for 7 generations,the yellow and black immature seed RNA bulked pools were constructed.SRAP was used to analysis the different cDNA pools transcribed from RNA pools.Totally,2 100 amplifiedbands were obtained with 996 pairs of SRAP primer and the repeated bands were 65.2%.Twelve steady differential fragments with length of 100-300 bp between yellow and black immature seeds were acquired.Seven stable differential fragments were sorted out,cloned and sequenced.Sequence analysis revealed that only two of the fragments had significant homologous nucleotide sequence with Arabidopsis thaliana,one was similar to NAD＋ADP-ribosyl transferase and the other was peptide transporter protein 3.The results indicated that cDNA pools with SRAP marker could be used for differential display on Brassica napus,and the two fragments may be related to the seed coat color expression of Brassica napus.
出处
《作物学报》
CAS
CSCD
北大核心
2008年第3期526-529,共4页
Acta Agronomica Sinica
基金
国家自然科学基金重点项目(30330400)
作者简介
马爱芬（1980-），女。河南清丰县人，硕士，研究方向：植物分子育种。E-mail：mafeifei3@163．com
通讯作者（Corresponding author）：刘列钊（1973-），男，四川泸州人，副教授，从事作物遗传育种。Tel：023-68250701；E-mail：liezhao2003@126．com
