生物技术通讯
生物技術通訊
생물기술통신
LETTERS IN BIOTECHNOLOGY
2014年
4期
467-473
,共7页
虞飞博%周洁%王栩鸣%杨勇%余初浪%程晔%严成其%陈剑平
虞飛博%週潔%王栩鳴%楊勇%餘初浪%程曄%嚴成其%陳劍平
우비박%주길%왕허명%양용%여초랑%정엽%엄성기%진검평
OsPR1b基因%启动子%表达特性%水稻
OsPR1b基因%啟動子%錶達特性%水稻
OsPR1b기인%계동자%표체특성%수도
OsPR1b gene%promoter%expression characteristic%rice
目的：分析水稻病程相关基因OsPR1b的表达特性，以进一步了解其表达和调控机制。方法：利用PCR技术从水稻日本晴基因组中扩增OsPR1b基因的启动子片段，命名为OsPR1bp，并构建相应的OsPR1bp：GUS融合表达载体，采用农杆菌介导的转基因技术获得转基因植株，进行GUS组织化学分析；利用Real-time PCR对OsPR1b基因在植物激素、非生物因子和水稻白叶枯菌（Xoo）毒性菌株P10（PXO124）处理下的表达水平进行分析。结果：GUS组织染色结果表明OsPR1b在水稻叶片中的表达量较高，而在茎、根、愈伤和花器中的表达量较低；植物激素水杨酸（SA）、茉莉酸甲酯（MeJA）、激动素（KT）、脱落酸（ABA）及NaCl、PEG均可不同程度地提高OsPR1b在叶片中的表达水平，Me-JA、KT和NaCl的处理能提高其在根部的表达水平，但这些激素在诱导OsPR1b在叶片和根部的表达程度上存在明显差异；单独接种Xoo毒性菌株P1024 h对OsPR1b表达的影响不大，而MeJA与其共同处理后则可显著增强其在叶片中的表达。结论：作为一种防卫基因，OsPR1b在健康植株中的表达水平较低，容易受盐/干旱胁迫及Xoo病原菌的诱导，多种植物激素如JA、KT和ABA很可能作为信号分子参与激活和介导了这种系统性的反应。
目的：分析水稻病程相關基因OsPR1b的錶達特性，以進一步瞭解其錶達和調控機製。方法：利用PCR技術從水稻日本晴基因組中擴增OsPR1b基因的啟動子片段，命名為OsPR1bp，併構建相應的OsPR1bp：GUS融閤錶達載體，採用農桿菌介導的轉基因技術穫得轉基因植株，進行GUS組織化學分析；利用Real-time PCR對OsPR1b基因在植物激素、非生物因子和水稻白葉枯菌（Xoo）毒性菌株P10（PXO124）處理下的錶達水平進行分析。結果：GUS組織染色結果錶明OsPR1b在水稻葉片中的錶達量較高，而在莖、根、愈傷和花器中的錶達量較低；植物激素水楊痠（SA）、茉莉痠甲酯（MeJA）、激動素（KT）、脫落痠（ABA）及NaCl、PEG均可不同程度地提高OsPR1b在葉片中的錶達水平，Me-JA、KT和NaCl的處理能提高其在根部的錶達水平，但這些激素在誘導OsPR1b在葉片和根部的錶達程度上存在明顯差異；單獨接種Xoo毒性菌株P1024 h對OsPR1b錶達的影響不大，而MeJA與其共同處理後則可顯著增彊其在葉片中的錶達。結論：作為一種防衛基因，OsPR1b在健康植株中的錶達水平較低，容易受鹽/榦旱脅迫及Xoo病原菌的誘導，多種植物激素如JA、KT和ABA很可能作為信號分子參與激活和介導瞭這種繫統性的反應。
목적：분석수도병정상관기인OsPR1b적표체특성，이진일보료해기표체화조공궤제。방법：이용PCR기술종수도일본청기인조중확증OsPR1b기인적계동자편단，명명위OsPR1bp，병구건상응적OsPR1bp：GUS융합표체재체，채용농간균개도적전기인기술획득전기인식주，진행GUS조직화학분석；이용Real-time PCR대OsPR1b기인재식물격소、비생물인자화수도백협고균（Xoo）독성균주P10（PXO124）처리하적표체수평진행분석。결과：GUS조직염색결과표명OsPR1b재수도협편중적표체량교고，이재경、근、유상화화기중적표체량교저；식물격소수양산（SA）、말리산갑지（MeJA）、격동소（KT）、탈락산（ABA）급NaCl、PEG균가불동정도지제고OsPR1b재협편중적표체수평，Me-JA、KT화NaCl적처리능제고기재근부적표체수평，단저사격소재유도OsPR1b재협편화근부적표체정도상존재명현차이；단독접충Xoo독성균주P1024 h대OsPR1b표체적영향불대，이MeJA여기공동처리후칙가현저증강기재협편중적표체。결론：작위일충방위기인，OsPR1b재건강식주중적표체수평교저，용역수염/간한협박급Xoo병원균적유도，다충식물격소여JA、KT화ABA흔가능작위신호분자삼여격활화개도료저충계통성적반응。
Objective: In order to gain further insight into the expression/regulation of marker gene OsPR1b in rice immune response, expression analysis of OsPR1b in rice was carried out. Methods: OsPR1b gene promoter fragment was amplified using the technology of PCR from rice Nipponbare genome and named OsPR1bp. The plas-mid of OsPR1bp::GUS was introduced into japonica rice via Agrobacterium-mediated transformation for further histo-chemical GUS analysis. In addition, the expression level of OsPR1b gene in rice leaves and roots was analyzed af-ter treated with plant hormones, abiotic factors and Xanthomonas oryzae pv. oryzae(Xoo) P10(PXO124) strain re-spectively. Results: The expression level of OsPR1b gene was much higher in leaf than that in stem, root, callus and flower. The expression of OsPR1b in leaves was differentially increased under the treatments of salicylic acid (SA), methyl jasmonate(MeJA), kinetin(KT), abscisic acid(ABA) as well as NaCl and PEG. The expression of OsPR1b in roots was increased under the treatments of MeJA, KT and NaCl. However, the increased fold of ex-pression were different between leaves and roots. Inoculation with P10 alone for 24 h had no effect on OsPR1b ex-pression, while co-treated with MeJA can significantly enhanced the expression level of OsPR1b in leaves. Conclu-sion: OsPR1b is described as a defense gene and the promoter we isolated directed a very low GUS gene expres-sion under normal conditions but responded to salt/drought stress and Xoo, and plants hormones such as JA, KT and ABA probably act as signals to trigger and mediate the systematic response.
