Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (15): 3172-3179.doi: 10.3864/j.issn.0578-1752.2013.15.011

• HORTICULTURE • Previous Articles     Next Articles

Differential Expression Genes of ‘Dangshansuli’ Pear Russet Mutant Screened with DDRT-PCR

 HENG  Wei, CHENG  Xiu-Yun, JIA  Bing, LIU  Pu, LIU  Li, YE  Zhen-Feng, ZHU  Li-Wu   

  1. College of Horticulture, The Key Laboratory of Pomology, Anhui Agricultural University, Hefei 230036
  • Received:2012-12-25 Online:2013-08-01 Published:2013-05-17

PDF

534

Abstract: 【Objective】The objective of this experiment is to study the formation mechanism of ‘Dangshansuli’ pear russet mutant. 【Method】 The peels of ‘Dangshansuli’ pear and its mutant ‘Xiusu’ on 110 d after full bloom were used to screen the differential expression genes with mRNA differential display reverse transcription-polymerase chain reaction (DDRT-PCR). 【Result】 Based on the function annotation, the key differential genes related with russet formation were confirmed with real-time PCR on 90, 110, 120, 140 and 150 d. The results showed that 63 differential genes were screened with 60 high quality and 3 low quality genes, including alpha-tubulin, N-methyltransferase, mannose-6-phosphate isomerase-like, calmodulin-binding protein kinase, vacuolar H+-PPase and so on. The expressions of α-tubulin gene in the pericarp of ‘Xiusu’ were higher than those in ‘Dangshansuli’ pear at different stages, and expecially on the 90 d and 110 d after full bloom with 5 times and 3 times, respectively. On the 90 d and 110 d after full bloom, the expression of calcium/calmodulin-dependent serine/threonine protein kinase gene in the pericarp of ‘Xiusu’ were slightly higher than those in ‘Dangshansuli’ pear, while the expressions of Ca2+/CaM gene in the pericarp of ‘Xiusu’ were distinctly lower than those in ‘Dangshansuli’ pear during 120-150 d after full bloom. The expression of mannose-6-phosphate isomerase gene in the pericarp of ‘Dangshansuli’ pear and ‘Xiusu’ showed no distinct difference except 130 d after full bloom. 【Conclusion】 These results indicated that the pericarp of russet mutant was lignified with α-tubulin accumulation regulated with the expression of calcium/calmodulin-dependent serine/threonine protein kinase gene.

Key words: Pyrus , russet mutant , fruit peel , differential gene , real-time PCR

[1]曹玉芬. 我国红皮梨种质资源. 中国种业, 2001(1): 26-27.

Cao Y F. Germplasm resources of Chinese red peel pear. China Seeds, 2001(1): 26-27. (in Chinese)

[2]滕元文, 沈玉英, 周先章. 砂梨果皮锈斑成因及解决对策. 中国南方果树, 2005(34): 30-32.

Teng Y W, Shen Y Y, Zhou X Z. Rust spot of Pyrus pyrifolia Nakai: cause and control. South China Fruit, 2005(34): 30-32. (in Chinese)

[3]Kim D, Hwang J H, Shin Y U, Shin I S, Lee H J, Hong S S, Kang S J. Development of molecular markers linked to several fruit traits in orental pear. Acta Horticulturae, 2005, 671: 315-321.

[4]Inoue E, Kasumi M, Sakuma F, Anzai H, Amano K, and Hara H. Identification of RAPD marker linked to fruit skin color in Japanese pear (Pyrus pyrifolia Nakai). Scientia Horticulturae, 2006, 107(3): 254-258.

[5]宋伟, 王彩虹, 田义轲, 田伟, 殷豪. 梨果实褐皮性状的SSR标记. 园艺学报, 2010, 37(8): 1325-1328.

Song W, Wang  C H, Tian Y K, Tian W, Yin H. SSR molecular markers linked to the fruit russet skin of pear. Acta Horticulturae Sinica, 2010, 37(8): 1325-1328. (in Chinese)

[6]Lang P, Zhang C K，Ebel R C, Dane F, Dozier W A. Identification of cold acclimated genes in leaves of Citrus unshiu by mRNA differential display. Gene, 2005, 359: 111-118.

[7]刘凯, 曾继吾, 夏瑞, 陈金印, 易干军, 黄永红, 周碧容. 柑橘mRNA差异显示技术体系建立. 基因组学与应用生物学, 2009, 28(2): 339-344.

Liu K, Zeng J W, Xia R, Chen J Y, Yi G J, Huang Y H, Zhou B R. Establishing the system of Citrus mRNA differential display technology. Genomics and Applied Biology, 2009, 28(2): 339-344. (in Chinese)

[8]Hamano M, Miura H, Tabei Y. Analysis of differentially expressed mRNAs during the early developmental stage of strawberry (Fragaria ananassa Duch.) fruit by differential display technique. Journal of the Japanese Society for Horticulture Science, 1998, 67(2): 559-561.

[9]张今今, 王跃进. mRNA差异显示技术克隆中国葡萄属野生种核糖体RNA基因. 西北植物学报, 2005, 25(11): 2163-2167.

Zhang J J, Wang Y J. Cloning of ribosomal RNA gene in Chinese wild Vitis species by DDRT-PCR. Acta Botanica Boreali-Occidentalia Sinica, 2005, 25(11): 2163-2167. (in Chinese)

[10]李慧娥, 王西平, 王跃进, 杨亚州, 王倩. 中国野生毛葡萄钙调蛋白基因克隆及序列分析. 西北植物学报, 2010, 30(8): 1507-1513.

Li H E, Wang X P, Wang Y J, Yang Y Z, Wang Q. Isolation and sequence analysis of calmodulin gene of Chinese wild Vitis quinquangularis. Acta Botanica Boreali-Occidentalia Sinica, 2010, 30(8): 1507-1513. (in Chinese).

[11]王西平, 王倩, 王跃进. 华东葡萄抗白粉病基因差异表达cDNA克隆与序列分析. 西北农业学报, 2011, 20(9): 117-122.

Wang X P, Wang Q, Wang Y J. Cloning and analyzing the gene cDNA sequences of resistance to uncinula necator from Vitis pseudoreticulata. Acta Agriculturae Boreali-Occidentalia Sinica, 2011, 20(9): 117-122. (in Chinese)

[12]吴海波, 郑国华, 牛先前, 杜会香. DDRT-PCR技术分离枇杷幼果低温诱导相关基因. 热带作物学报，2010, 31(3): 416-421.

Wu H B, Zheng G H, Niu X Q, Du H X. Isolation of the cold stress genes in loquat fruit by DDRT-PCR technique. Chinese Journal of Tropical Crops, 2010, 31(3): 416-421. (in Chinese)

[13]杨转英, 胡桂兵, 王惠聪, 欧阳若, 陈厚彬. 不同着色期荔枝果皮总RNA的提取及mRNA差显分析. 果树学报, 2008, 25(2): 281-285.

Yang Z Y, Hu G B, Wang H C, Ouyang R, Chen H B. Total RNA extraction and mRNA differential display analysis of litchi pericarps at different coloring stages. Journal of Fruit Science, 2008, 25(2): 281-285. (in Chinese)

[14]李永强, 赵腮宝, 倪开诚, 陈文荣, 辛德东, 郭卫东. ‘短柄樱桃’花芽休眠解除过程中差异表达基因的研究. 园艺学报, 2012, 39(7): 1253-1262.

Li Y Q, Zhao S B, Ni K C, Chen W R, Xin D D, Guo W D. Studies on differentially expressed genes in flower bud during dormancy release in Prunus pseudocerasus Lindl. ‘Duanbing’. Acta Horticulturae Sinica, 2012, 39(7): 1253-1262. (in Chinese)

[15]张飞, 牛建新, 叶春秀, 刘娜, 高静涛, 朱天丁. 库尔勒香梨mRNA差异显示技术体系的建立. 石河子大学学报: 自然科学版, 2011, 29(2): 173-178.

Zhang F, Niu J X, Ye C X, Liu N, Gao J T, Zhu T D. Establishing the system of Korla fragrant pear mRNA differential display technology. Journal of Shihezi University: Natural Science, 2011, 29(2): 173-178. (in Chinese)

[16]王西成, 乔玉山, 欧春青, 王斐, 马力, 姜淑玲. 三种梨果皮总RNA提取方法的比较研究. 甘肃农业大学学报, 2010, 2: 91-94.

Wang X C, Qiao Y S, Ou C Q, Wang F, Ma L, Jiang S L. Extraction of total RNA from pericarp of pears with three methods. Journal of Gansu Agriculture University, 2010, 2: 91-94. (in Chinese)

[17]李晓峰, 李雪, 贾兵, 刘莉, 叶振风, 衡伟, 朱立武. ‘砀山酥梨’褐皮芽变木质素含量及相关酶活性与CCoAOMT表达量分析. 园艺学报, 2012, 39(5) : 828-836.

Li X F, Li X, Jia B, Liu L, Ye Z F, Heng W, Zhu L W. Analysis of enzyme activity and lignin content and expression of CCoAOMT gene in the pericarp of ‘Dangshansuli’ and its russet mutant. Acta Horticulturae Sinica, 2012, 39(5): 828- 836. (in Chinese)

[18]陈硕, 陈珈. 植物中钙依赖蛋白激酶（CDPKs）的结构与功能. 植物学通报, 2001, 18(2): 143-148.

Chen S, Chen J. The structure and function of calcium-dependent protein kinases in plant. Chinese Bulletin of Botany, 2001, 18(2): 143-148. (in Chinese)

[19]孙学辉, 路铁刚, 贾士荣, 黄大昉. 基因组水平上水稻受体激酶基因家族的生物信息学分析. 中国农业科学, 2004, 37(3): 322-327.

Sun X H, Lu T G, Jia S R, Huang D F. Genome-wide bioinformatics analysis of the rice receptor-like kinase family. Scientia Agricultura Sinica, 2004, 37(3): 322-327. (in Chinese)

[20]叶子, 黄聪聪, 于荣. 保卫细胞微管骨架参与蛋白丝氨酸/苏氨酸磷酸化调节的气孔运动. 中国农业科学, 2012, 45(21): 4351-4360.

Ye Z, Huang C C, Yu R. Microtubules in guard cells involve in serine/threonine phosphorylation during stomatal movement. Scientia Agricultura Sinica, 2012, 45(21): 4351-4360. (in Chinese)

[21]Asano T, Tanaka N, Yang G, Hayashi N, KomatsuS. Genome-wide identification of the rice calcium-dependent protein kinase and its closely related kinase gene families: comprehensive analysis of the CDPKs gene family in rice. Plant and Cell Physiology, 2005, 46(2): 356–366.

[22]粱小娥, 张大鹏, 贾文锁. 苹果和葡萄果实蛋白激酶特性分析. 植物生理学报, 2000, 26(3): 257-262.

Liang X E, Zhang D P, Jia W S. Analysis of protein kinase characteristics in developing apple and grape fruits. Acta Phytophysiologica Sinica, 2000, 26(3): 257-262. (in Chinese)

[23]郁松林，黄卫东. 热胁迫对葡萄叶片中蛋白激酶活性的影响. 中国农业科学, 2005, 38(1): 128-134.

Yu S L, Huang W D. Influence of heat stress on the activity of protein kinase in grape leaves. Scientia Agricultura Sinica, 2005, 38(1): 128-134. (in Chinese)

[24]杜中军, 王家保, 黄俊生, 翟衡, 徐兵强. 番木瓜丝氨酸/苏氨酸蛋白激酶类抗病基因同源序列的克隆与特征分析. 果树学报, 2006, 23(1): 46-50.

Du Z J, Wang J B, Huang J S, Zhai H, Xu B Q. Isolation and characterization of serine/threonine protein kinase-like disease resistance gene analogs from papaya (Carica papaya). Journal of Fruit Science, 2006, 23(1): 46-50. (in Chinese)

[25]张玄兵, 杜中军, 黄俊生, 王家保, 徐兵强. 编码杧果丝氨酸/苏氨酸蛋白激酶类蛋白抗病基因同源序列的克隆与分析. 华中农业大学学报, 2006, 25(2): 186-189.

Zhang X B, Du Z J, Huang J S, Wang J B, Xu B Q. Cloning and analysis of RGAs genes coding serine/threonine protein kinase-like proteins from Mango (Mangifera indica L.). Journal of Huazhong Agricultural University, 2006, 25(2): 186-189. (in Chinese)

[26]王萌, 王俊, 刘洪, 李静, 段可, 叶正文, 高清华. 草莓钙依赖蛋白激酶基因的克隆与表达分析. 西北植物学报, 2011, 31(5): 875-880.

Wang M, Wang J, Liu H, Li J, Duan K, Ye Z W, Gao Q H. Gene cloning and expression analysis of calcium-dependent protein kinase in strawberry. Acta Botanica Boreali-Occidentalia Sinica, 2011, 31(5): 875-880. (in Chinese)

[27]Yand G, Jan A Komatsu S. Characterization of β-tubulin 4 regulated by gibberellins in rice leaf sheath. Biologia Plantarum, 2009, 53(3): 422-428.
[1] ZHAI XiaoHu,LI LingXu,CHEN XiaoZhu,JIANG HuaiDe,HE WeiHua,YAO DaWei. Quantitative Detection Technology of Porcine-Derived Materials in Meat by Real-time PCR [J]. Scientia Agricultura Sinica, 2023, 56(1): 156-164.
[2] ZHU ChunYan,SONG JiaWei,BAI TianLiang,WANG Na,MA ShuaiGuo,PU ZhengFei,DONG Yan,LÜ JianDong,LI Jie,TIAN RongRong,LUO ChengKe,ZHANG YinXia,MA TianLi,LI PeiFu,TIAN Lei. Effects of NaCl Stress on the Chlorophyll Fluorescence Characteristics of Seedlings of Japonica Rice Germplasm with Different Salt Tolerances [J]. Scientia Agricultura Sinica, 2022, 55(13): 2509-2525.
[3] XU Chen,WANG WenJing,CAO Shan,LI RuXue,ZHANG BeiBei,SUN AiQing,ZHANG ChunQing. Mechanism of DA-6 Treatment Regulating Wheat Seed Vigor After Anthesis [J]. Scientia Agricultura Sinica, 2021, 54(9): 1821-1834.
[4] Tao WANG,Yu HAN,Li PAN,Bing WANG,MaoWen SUN,Yi WANG,YuZi LUO,HuaJi QIU,Yuan SUN. Development of a TaqMan Real-Time PCR Targeting the MGF360-13L Gene of African Swine Fever Virus [J]. Scientia Agricultura Sinica, 2021, 54(5): 1073-1080.
[5] ZHANG DaoWei,KANG Kui,YU YaYa,KUANG FuPing,PAN BiYing,CHEN Jing,TANG Bin. Characteristics and Immune Response of Prophenoloxidase Genes in Sogatella furcifera [J]. Scientia Agricultura Sinica, 2020, 53(15): 3108-3119.
[6] LI WenXue, XIAO RuiGang, LÜ MiaoMiao, DING Ning, SHI HuaRong, GU PeiWen. Establishment and Application of Real-Time PCR for Quantitatively Detecting Plasmopara viticola in Vitis vinifera [J]. Scientia Agricultura Sinica, 2019, 52(9): 1529-1540.
[7] BAI HuiYang, LU Geng, LU JunXing, GUAN Li, TANG Xin, ZHANG Tao. Cloning and Expression Analysis of Jasmonic Acid Carboxyl Methyltransferase Gene from Perilla frutescens [J]. Scientia Agricultura Sinica, 2019, 52(9): 1657-1666.
[8] DING YanJuan,LIU YongKang,LUO YuJia,DENG YingMei,XU HongXing,TANG Bin,XU CaiDi. Potential Functions of Nilaparvata lugens GSK-3 in Regulating Glycogen and Trehalose Metabolism [J]. Scientia Agricultura Sinica, 2019, 52(7): 1237-1246.
[9] TANG Bin,SHEN QiDa,ZENG BoPing,XIAO ZhongJiu,QIU LingYu,PAN BiYing,LI Kun,ZHANG DaoWei. Characteristics, Developmental Expression and RNAi Effect Analysis of a Novel Trehalose-6-Phosphate Synthase Gene in Nilaparvata lugens [J]. Scientia Agricultura Sinica, 2019, 52(3): 466-477.
[10] ZHANG Fei,WANG YanQiu,ZHU Kai,ZHANG ZhiPeng,ZHU ZhenXing,LU Feng,ZOU JianQiu. Comparative Transcriptome Analysis of Different Salt Tolerance Sorghum (Sorghum bicolor L. Moench) Under Salt Stress [J]. Scientia Agricultura Sinica, 2019, 52(22): 4002-4015.
[11] BAI Hui, SONG ZhenJun, WANG YongFang, QUAN JianZhang, MA JiFang, LIU Lei, LI ZhiYong, DONG ZhiPing. Identification and Expression Analysis of MYB Transcription Factors Related to Rust Resistance in Foxtail Millet [J]. Scientia Agricultura Sinica, 2019, 52(22): 4016-4026.
[12] LIU FanQi,WAN GuiJun,ZENG LuYing,LI ChunXu,PAN WeiDong,CHEN FaJun. Selection of Stable Internal Reference Genes for Transcript Expression Analyses in Laodelphax striatellus Under Near-Zero Magnetic Field [J]. Scientia Agricultura Sinica, 2019, 52(19): 3346-3356.
[13] ZHANG DaoWei,YU YaYa,PAN BiYing,KANG Kui,ZENG BoPing,CHEN Jing,TANG Bin. Regulation Function of Trehalose-6-phosphate Synthase Genes on Chitin Synthesis in Sogatella furcifera [J]. Scientia Agricultura Sinica, 2019, 52(19): 3357-3366.
[14] XIE Jie,WANG Ming,DING HongYing,LI Qing,WANG WanXing,XIONG XingYao,QIN YuZhi. Expression and Structural Analysis of SC MI390-5p and Its Target Genes in Potato Response to Low Temperature [J]. Scientia Agricultura Sinica, 2019, 52(13): 2295-2308.
[15] ZHANG ShuangNa, LI ZhengNan, FAN XuDong, ZHANG ZunPing, REN Fang, HU GuoJun, DONG YaFeng. Establishment of RT-LAMP Assay for Detection of Apple chlorotic leaf spot virus (ACLSV) [J]. Scientia Agricultura Sinica, 2018, 51(9): 1706-1716.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备09089781号-30
Copyright 2018 © Editorial office of Scientia Agricultura Sinica
Tel: 86-010-82106279    E-mail: zgnykx@mail.caas.net.cn
Supported by Beijing Magtech Co.Ltd