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Cloning, expression analysis, and detection method of the S12-RNase gene existing in three pear species (Pyrus bretschneideri, P. pyrifolia, and P. ussuriensis)
Zhang Lin 1 * #,Liu Min 2,Tan Xiaofeng 2,Song Zhibo 2,Long Hongxu 2,Cao Yufen 3,Li Xiugen 4
1.Key Laboratory of Cultivation and Protection for Non-Wood Forest Trees, Ministry of Education, Central South University of Forestry and Technology, ChangSha 410004
2.Key Laboratory of Cultivation and Protection for Non-Wood Forest Trees, Ministry of Education, Central South University of Forestry and Technology, Changsha 410004, China
3.Research Institute of Pomology, Chinese Academy of Agricultural Sciences, Xingcheng 125100, China
4.Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences, Zhengzhou 450009, China
*Correspondence author
#Submitted by
Subject:
Funding: Senior Talent Introduction Foundation of Central South University of Forestry & Technology (No.104-0129), Scientific Research Foundation of Central South University of Forestry & Technology (No.2009004A), New Teachers‘ Foundation for Doctor Stations, Ministry of Education(No.20094321120001)
Opened online:30 November 2012
Accepted by: none
Citation: Zhang Lin,Liu Min,Tan Xiaofeng.Cloning, expression analysis, and detection method of the S12-RNase gene existing in three pear species (Pyrus bretschneideri, P. pyrifolia, and P. ussuriensis)[OL]. [30 November 2012] http://en.paper.edu.cn/en_releasepaper/content/4497964
 
 
A specific forward primer was designed based on the principle of highest similarity, and employed in 3' RACE using Chinese white pear (Pyrus bretschneideri) cultivar 'Yingzhiqing' as materials. The full length cDNA of PbS12-RNase was successfully amplified and deposited in GenBank (Accession No. EU081889). At the amino acid level, the PbS12-RNase exhibited the highest similarity (97.3%) with MdSf-RNase of Malus domestica, and only six amino acid differences were present in the two S-RNases. Phylogenetic analysis of rocaceous S-RNases indicated that the PbS12-RNase clustered with maloideous S-RNases, forming a subfamily-specific not species-specific group. Some intraspecific genetic distance of the S-RNases, in addition, was greater than interspecific genetic distance. In 'Yingzhiqing', the PbS12-allele was specifically expressed in style. Moreover, the expression level of this gene was extremely low at the small bud stage, and subsequently increased rapidly at the bid bud stage and bell stage. A method for rapid detection of the PbS12-allele was developed via PbS12-allele-specefic primers design based on multiple sequence comparisons. Application of the PbS12-allele-specefic primers in 59 cultivars from four pear species showed that the PbS12-allele not only existed in P. bretschneideri, but in P. pyrifolia and P. ussuriensis as well. The present study could provide a scientific base for fully clarifying the mechanism of pear GSI at the molecular level.
Keywords:gametophytic self-incompatibility; reverse transcript-PCR; Rapid Amplification of cDNA Ends; S-RNase; phylogenetic tree; spatio-temporal expression patterns
 
 
 

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