江苏农业学报
江囌農業學報
강소농업학보
JIANGSU JOURNAL OF AGRICULTURAL SCIENCES
2014年
6期
1464-1471
,共8页
王宏%蔺经%李晓刚%杨青松%常有宏
王宏%藺經%李曉剛%楊青鬆%常有宏
왕굉%린경%리효강%양청송%상유굉
杜梨%蛋白磷酸酶PP2C%基因克隆%表达分析
杜梨%蛋白燐痠酶PP2C%基因剋隆%錶達分析
두리%단백린산매PP2C%기인극륭%표체분석
Pyrus betulaefolia Bunge%protein phosphatase PP2C%gene c1oning%expression ana1ysis
2C型丝氨酸/苏氨酸蛋白磷酸酶(PP2C)是植物体内脱落酸ABA信号传导关键负调控酶,在植物耐非生物胁迫中发挥着重要的作用。本研究利用RT-PCR技术,克隆出编码杜梨PP2C蛋白的基因PbPP2C,其核苷酸序列全长1353 bp,开放阅读框长1263 bp,编码422个氨基酸残基,这些残基中包括了与二价金属离子结合的MED、DGH、DG和D结构域。系统发生分析结果表明,该氨基酸与拟南芥AHG3亲缘关系最近。实时荧光定量PCR分析结果表明:盐胁迫条件下,PbPP2C基因在根、茎、叶中的表达12 h达到峰值,同时在根中72 h达到第二个峰值;干旱胁迫条件下,PbPP2C基因在根、茎、叶中表达72 h达到峰值。说明PbPP2C与杜梨耐盐和耐干旱胁迫存在紧密关联。
2C型絲氨痠/囌氨痠蛋白燐痠酶(PP2C)是植物體內脫落痠ABA信號傳導關鍵負調控酶,在植物耐非生物脅迫中髮揮著重要的作用。本研究利用RT-PCR技術,剋隆齣編碼杜梨PP2C蛋白的基因PbPP2C,其覈苷痠序列全長1353 bp,開放閱讀框長1263 bp,編碼422箇氨基痠殘基,這些殘基中包括瞭與二價金屬離子結閤的MED、DGH、DG和D結構域。繫統髮生分析結果錶明,該氨基痠與擬南芥AHG3親緣關繫最近。實時熒光定量PCR分析結果錶明:鹽脅迫條件下,PbPP2C基因在根、莖、葉中的錶達12 h達到峰值,同時在根中72 h達到第二箇峰值;榦旱脅迫條件下,PbPP2C基因在根、莖、葉中錶達72 h達到峰值。說明PbPP2C與杜梨耐鹽和耐榦旱脅迫存在緊密關聯。
2C형사안산/소안산단백린산매(PP2C)시식물체내탈락산ABA신호전도관건부조공매,재식물내비생물협박중발휘착중요적작용。본연구이용RT-PCR기술,극륭출편마두리PP2C단백적기인PbPP2C,기핵감산서렬전장1353 bp,개방열독광장1263 bp,편마422개안기산잔기,저사잔기중포괄료여이개금속리자결합적MED、DGH、DG화D결구역。계통발생분석결과표명,해안기산여의남개AHG3친연관계최근。실시형광정량PCR분석결과표명:염협박조건하,PbPP2C기인재근、경、협중적표체12 h체도봉치,동시재근중72 h체도제이개봉치;간한협박조건하,PbPP2C기인재근、경、협중표체72 h체도봉치。설명PbPP2C여두리내염화내간한협박존재긴밀관련。
Protein phosphatase PP2C is an important negative regu1ator and exerts crucia1 ro1es when p1ants are in abiotic stress. The cDNA of PbPP2C encoding protein phosphatase PP2C was c1oned by RT-PCR from sa1t-to1erant Pyrus betulaefolia Bunge. The 1ength of nuc1eotide sequence was 1 353 bp in 1ength with a 1 263 bp ORF ( open reading frame) , encoding 422 amino acids, which inc1uded meta1 ions-binding domain such as MED, DGH, DG and D amino acid resi-dues. Phy1ogenetic tree showed that PbPP2C was c1ustered c1ose1y with AHG3 of Arabidopsis. The resu1ts by qRT-PCR sug-gested that the expression of PbPP2C in roots, tems and 1eaf of pear achieved the peak at 12-h sa1t stress and the second peak at 72 h in roots. Under drought stress, the expression of PbPP2C in roots, stems and 1eaf of pear reached the peak at 72 h. It is suggested that the PbPP2C gene is c1ose1y re1ated to sa1t to1erance and drought to1erance of Pyrus betulaefolia Bunge.