Plant Mol Biol Rep (2011)29:723-732 D0I10.1007s11105-010-0285-y A cDNA Clone of BcHSP81-4 from the Sterility Line (Pol CMS)of Non-heading Chinese Cabbage (Brassica campestris ssp.chinensis) Tongkun Liu·Xilin Hou·Jingyi Zhang·Yuping Song" Shuning Zhang·Ying Li Published online:8 January 2011 C Springer-Verlag 2011 Abstract BcHSP81-4 gene,a member of heat shock Introduction proteins,was identified from a suppression subtractive hybridization cDNA library in non-heading Chinese cab- During their lifetime,plant species can be subjected to bage (Brassica campestris ssp.chinensis Makino).The various stressful environments to which they respond and deduced amino acid sequence of the BcHSP81-4 cDNA adapt by means of physiological,developmental,and revealed that it has high homology to other plant organelle biochemical changes.One of the most thoroughly charac- isoforms and similar homology to both cytoplasmic and terized is the induction of heat shock proteins (HSPs). prokaryotic HSP90s.To study the regulation of gene which are a group of universal and evolutionarily conserved expression,BcHSP81-4 genes in maintainer and sterility protein (Li et al.2009).Generally,HSPs are grouped into lines were monitored at different development stages and at five major families on the basis of their molecular size in different stress treatments.Real-time PCR was used for kilodaltons:HSP110,HSP90,HSP70,HSP60,and the low quantification of BcHSP8/-4 mRNA.These results indicate molecular weight HSP families (Scharf et al.2001).The that BcHSP81-4 is not responsive to heat shock at least at heat shock proteins contribute to various cellular processes 35C,while it is very responsive to salt and cold stress. including signal transduction and the folding,localization, And high expression of BcHSP81-4 in the bud of sterile accumulation,and degradation of protein molecules in both line suggests that it may play prominent roles in sterility of plant and animal species (Yu et al.2005). pol CMS in non-heading Chinese cabbage. Several members of the plant HSP gene family have been isolated.Plants contain a wide array of HSPs.HSPs Keywords CMS.Heat shock protein.Non-heading can be induced by a variety of environmental factors Chinese cabbage.Real-time PCR including heat(Piano et al.2005),low temperature (Taylor et al.2005),and other environmental stresses such as Abbreviations drought (Vasquez-Robinet et al.2008;Xu et al.2009), CMS Cytoplasmic male sterility salinity or flooding (Liu et al.2006;Lin et al.2010), EST Expressed sequence tag cadmium or arsenite (Milioni and Hatzopoulos 1997), HSP Heat shock protein ultraviolet light,oxidative stress(Song et al.2009),osmotic SSH Suppression subtractive hybridization stress,and pathogen infection (Swindell et al.2007). However,HSPs are not only present after stress stimulation. The same proteins have been shown to be essential components of cells and developmental processes under normal physiolog- T.Liu·X.Hou·J.Zhang·Y.Song·S.Zhang·Y.Li(☒) ical conditions (Rutherford and Lindquist 1998;Fu et al. State Key Laboratory of Crop Genetics and Germplasm 2010).For example,HSP82 gene shows strong heat Enhancement,Key Laboratory of Southern Vegetable Crop inducibility at heat shock temperatures,while the HSP81 Genetic Improvement,Ministry of Agriculture, Nanjing Agricultural University, gene is strongly expressed in the absence of heat shock during Nanjing 210095,People's Republic of China the pre-meiotic and meiotic prophase stages of pollen e-mail:yingli@njau.edu.cn development but is only mildly heat inducible in leaf tissue ②SpringerA cDNA Clone of BcHSP81-4 from the Sterility Line (Pol CMS) of Non-heading Chinese Cabbage (Brassica campestris ssp. chinensis) Tongkun Liu & Xilin Hou & Jingyi Zhang & Yuping Song & Shuning Zhang & Ying Li Published online: 8 January 2011 # Springer-Verlag 2011 Abstract BcHSP81-4 gene, a member of heat shock proteins, was identified from a suppression subtractive hybridization cDNA library in non-heading Chinese cab￾bage (Brassica campestris ssp. chinensis Makino). The deduced amino acid sequence of the BcHSP81-4 cDNA revealed that it has high homology to other plant organelle isoforms and similar homology to both cytoplasmic and prokaryotic HSP90s. To study the regulation of gene expression, BcHSP81-4 genes in maintainer and sterility lines were monitored at different development stages and at different stress treatments. Real-time PCR was used for quantification of BcHSP81-4 mRNA. These results indicate that BcHSP81-4 is not responsive to heat shock at least at 35°C, while it is very responsive to salt and cold stress. And high expression of BcHSP81-4 in the bud of sterile line suggests that it may play prominent roles in sterility of pol CMS in non-heading Chinese cabbage. Keywords CMS . Heat shock protein . Non-heading Chinese cabbage . Real-time PCR Abbreviations CMS Cytoplasmic male sterility EST Expressed sequence tag HSP Heat shock protein SSH Suppression subtractive hybridization Introduction During their lifetime, plant species can be subjected to various stressful environments to which they respond and adapt by means of physiological, developmental, and biochemical changes. One of the most thoroughly charac￾terized is the induction of heat shock proteins (HSPs), which are a group of universal and evolutionarily conserved protein (Li et al. 2009). Generally, HSPs are grouped into five major families on the basis of their molecular size in kilodaltons: HSP110, HSP90, HSP70, HSP60, and the low molecular weight HSP families (Scharf et al. 2001). The heat shock proteins contribute to various cellular processes including signal transduction and the folding, localization, accumulation, and degradation of protein molecules in both plant and animal species (Yu et al. 2005). Several members of the plant HSP gene family have been isolated. Plants contain a wide array of HSPs. HSPs can be induced by a variety of environmental factors including heat (Piano et al. 2005), low temperature (Taylor et al. 2005), and other environmental stresses such as drought (Vasquez-Robinet et al. 2008; Xu et al. 2009), salinity or flooding (Liu et al. 2006; Lin et al. 2010), cadmium or arsenite (Milioni and Hatzopoulos 1997), ultraviolet light, oxidative stress (Song et al. 2009), osmotic stress, and pathogen infection (Swindell et al. 2007). However, HSPs are not only present after stress stimulation. The same proteins have been shown to be essential components of cells and developmental processes under normal physiolog￾ical conditions (Rutherford and Lindquist 1998; Fu et al. 2010). For example, HSP82 gene shows strong heat inducibility at heat shock temperatures, while the HSP81 gene is strongly expressed in the absence of heat shock during the pre-meiotic and meiotic prophase stages of pollen development but is only mildly heat inducible in leaf tissue T. Liu : X. Hou : J. Zhang : Y. Song : S. Zhang : Y. Li (*) State Key Laboratory of Crop Genetics and Germplasm Enhancement, Key Laboratory of Southern Vegetable Crop Genetic Improvement, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, People’s Republic of China e-mail: yingli@njau.edu.cn Plant Mol Biol Rep (2011) 29:723–732 DOI 10.1007/s11105-010-0285-y