河北杨脱水素基因(Phdhn1)的克隆与胁迫表达分析

为了鉴定河北杨中的脱水素基因并研究其在逆境胁迫中的表达规律,以初步分析脱水素基因在河北杨逆境胁迫中的调控功能,在实验室前期工作的基础上设计引物,通过PCR方法扩增河北杨cDNA文库获取脱水素基因,并通过半定量RT-PCR方法研究其在逆境中的表达情况。结果表明,河北杨脱水素全长基因856 bp,共编码227个氨基酸。同源性分析表明,河北杨中的脱水素基因与已知的其他杨树脱水素高度同源,因此将该基因定名为Phdhn1。半定量RT-PCR结果表明在干旱、低温、高温、高盐及ABA溶液不同时间段(0、0.5、1、2、5、10、24 h)的胁迫下,河北杨Phdhn1在干旱、ABA以及低温胁迫下均出现表达差异。该研究初步阐述了Phdhn1的逆境表达模式,说明了Phdhn1在河北杨逆境胁迫中起到的重要作用。     

低温促进白菜开花的效应分析

为了研究低温对白菜开花的效应,以普通白菜品种杭州油冬儿为试材,研究了萌动种子和幼苗低温处理对白菜开花及处理过程中可溶性糖、淀粉、可溶性蛋白质和内源激素含量的变化.结果表明,无论萌动种子还是幼苗低温处理都促进白菜植株开花;处理过程中,萌动种子与幼苗植株叶片的可溶性糖含量均升高,淀粉和可溶性蛋白质含量均下降,总的C/N值升高;萌动种子低温处理后植株的GA_3、IAA含量升高,而ABA含量下降,同时GA/ABA、IAA/ABA的值也升高,而GA/IAA的值却先下降后上升;幼苗低温处理使植株的GA_3、IAA和ABA三者的含量均升高,而且GA/ABA、GA_3/IAA,IAA/ABA的值也都升高.说明GA_3,IAA相对含量的增加有利于白菜的开花.     

脱落酸与赤霉素对瓜尔豆蔗糖代谢酶类的影响

研究了脱落酸与赤霉素对瓜尔豆叶片和种子发育过程中糖代谢相关酶活性的影响。结果表明,脱落酸与赤霉素处理均能降低酸性转化酶活性,赤霉素(500mg/L)对中性转化酶与酸性转化酶活性降低程度最大。赤霉素(500mg/L及750mg/L)在整个生育期增加叶片蔗糖合成酶与蔗糖磷酸合成酶活性,脱落酸(2.64mg/L,1.32mg/L及3.96mg/L)及赤霉素(250mg/L)均不同程度地降低蔗糖合成酶与蔗糖磷酸合成酶活性。各处理间差异达到显著水平。低浓度的脱落酸(1.32mg/L)降低种子中性转化酶和蔗糖磷酸合成酶活性;高浓度的脱落酸(2.64mg/L及3.96mg/L)能提高种子的中性转化酶、酸性转化酶、蔗糖合成酶与蔗糖磷酸合成酶活性。低浓度的赤霉素(250mg/L)能相对降低瓜尔豆种子内的中性转化酶、酸性转化酶、蔗糖合成酶与蔗糖磷酸合成酶活性,而高浓度的赤霉素(500mg/L及750mg/L)则明显地升高中性转化酶、酸性转化酶、蔗糖合成酶与蔗糖磷酸合成酶活性,表现出较强的蔗糖合成与分解能力。 ;     

花生种子休眠解除过程中相关基因的表达分析

种子休眠性是花生重要的农艺性状,外源乙烯利能诱导花生种子休眠的解除,为了阐明乙烯利作用下花生种子休眠解除的分子机制,设置吸胀的休眠种子为对照,100 mg L–1乙烯利处理吸胀休眠种子后不同时间的样品(AE1、AE2、AE3)进行转录组分析,比较了花生种子休眠解除过程中ABA、GA、ETH、auxin相关基因的表达。结果表明,15个与GA、40个与ABA、60个与ETH、56个与auxin相关的unigenes在花生种子休眠解除过程中表现显著差异表达。荧光定量PCR结果显示,ABA合成关键基因Ah NCED2和代谢关键基因Ah CYP707A1在种子休眠解除过程中均受外源乙烯利诱导,表达差异显著;在休眠和无休眠种子吸胀萌发过程中,Ah NCED2和Ah CYP707A1的表达趋势不同,Ah NCED2对于种子休眠的维持发挥积极作用,而Ah CYP707A1对于种子休眠解除发挥积极作用。     

Effect of genetic background on the target fatty acids of acyl-ACP thioesterase transgenes in Brassica napus

Acyl‐acyl carrier protein (ACP) thioesterase (TE) is involved in the biosynthetic fatty acid pathway of plants. Conventional canola lines transformed individually with the bay‐TE (Uc FatB1), elm‐TE (Ua FatB1), nutmeg‐TE (Mf FatB1) or Cuphea‐TE transgene (Ch FatB1), produce seed oil with modified fatty acid compositions. This study assessed the effects of genetic background, cytoplasm, maternal parent, and interaction of different TE transgenes, on the target fatty acids using F1 seeds and double haploid (DH) lines. The F₁ seeds were produced by crossing four TE transgenic parental lines and three non‐transgenic cultivars with distinct fatty acid compositions. The DH lines were developed from microspores of F₁ plants. DH lines from different crosses showed that genetic background does not have an effect on the relative levels of the target fatty acids of the same TE, indicating the stability of the substrate specificity of the TE within canola. However, significant effects of genetic background on the content of the major target fatty acids, lauric acid (C12:0) or palmitic acid (C16:0) depending on the TE, were observed. Expression of the TE in low erucic acid (C22:1) genotypes resulted in higher target fatty acid levels than expression in high C22:1 genotypes. Reciprocal crosses showed maternal effects, but not cytoplasmic effects. In addition, co‐expression of two different TE transgenes in the same seeds was observed. These results indicate the importance of selection for appropriate genetic backgrounds in order to maximize the expression of the target fatty acids of TE transgenes, and also indicate potential uses of TE co‐expression in modifying canola seed oil.     

Effect of light on endogenous levels of gibberellin and abscisic acid in seed germination of photoblastic weedy rice (Oryza sativa L.)

The effect of red (R) and R/far-red (FR) lights on endogenous gibberellin (GA) and abscisic acid (ABA) content was first investigated during the germination of photoblastic black-hulled weedy rice (PBWR) seeds. The R light-treated PBWR seeds germinated after 36–48 h and germination was increased to 63% at 72 h. However, the FR light-treated seeds after R light treatment, suppressed the R light effect showing only 11% germination even at 72 h after the light treatment. The PBWR seed treated with R light rapidly increased the endogenous level of GA₁ to about 200 times at 12 h before seed germination as compared with R/FR (control) which suppressed the effect of R light. The contents of other GAs like GA₁₂, GA₅₃, GA₁₉, GA₂₀, and GA₈ were not affected by the R light irradiation. These results showed that the major biosynthetic pathway of GAs in PBWR seeds is the early 13-hydroxylation pathway leading to GA₁, which was suggested to be physiologically active in the PBWR seed germination. The decrease in the level of ABA in the R light-treated seeds was greater than the R/FR light-treated seeds, indicating that the balance of endogenous GA1 and ABA is responsible for the induction of germination in the PBWR seed.     

Response of Wheat Grain to ABA and Imbibition at Low Temperature

During after-ripening of wheat grains, embryo responsiveness to abscisic acid (ABA) decreases in parallel with the loss of grain dormancy. Dormant grains of Triticum aestivum L. line ‘Kitakei-1354’ that had lost almost half their dormancy due to chilling still respond to ABA like fully dormant grains. Imbibition a: low temperature appears to break the dormancy without change of embryo responsiveness to ABA. Gibberellin (GA) and ABA-regulated α-amylase synthesis of the embryos. This is amplified when the grains are imbibed at low temperature. Imbibition at low temperature appears to condition cells and tissues of the embryo for germination and α-amylase synthesis.     

葡萄VvMSA基因的克隆及表达特性分析

为了解VvMSA基因的功能,以抗性葡萄品种Vidal Blanc组培苗为试材,克隆得到VvMSA基因,对其序列进行了生物信息学分析,并利用实时定量PCR分析了其组织表达特性和多种非生物胁迫和信号应答表达特性。结果表明,VvMSA基因片段大小为450 bp,编码149个氨基酸序列。生物信息学分析结果显示,VvMSA蛋白分子量约为16.703 k Da,等电点为5.68,不稳定系数为41.71,推测为不稳定蛋白。VvMSA含有65个氨基酸组成的ABA/WDS保守结构域。在进化上属于单独一个分支,它和已报道过的番茄Le ASR1分别属于同一个祖先进化出的2个分支。实时荧光定量PCR分析显示,VvMSA在葡萄不同组织中均有表达,花中表达量最高。多种逆境胁迫因子如盐、干旱和低温等能诱导VvMSA不同程度的上调表达,盐胁迫3 h时诱导表达量最高。同时,VvMSA受逆境胁迫信号分子NO和H_2S不同程度诱导表达上调,而SA则抑制VvMSA表达。激素ABA、GA_3、IAA和ET均能不同程度诱导VvMSA表达上调,并且VvMSA盐处理下的表达模式与ABA诱导的类似。综合以上结果推断VvMSA可能通过调控ABA信号途径来调节葡萄对盐胁迫应答。     

外源ABA抑制水稻种子发芽的生理机制

比较研究了外源植物激素ABA（脱落酸）与植物生长抑制剂MH（马来酰肼）对水稻种子发芽的抑制作用,以期阐明ABA抑制水稻种子发芽的生理学作用与机制。研究结果表明ABA与MH虽然都具有抑制水稻种子发芽的作用,但两者生理作用性质不同,ABA抑制水稻种子发芽是延缓种子发芽,而MH是降低种子活性,对种子发芽具遏制、伤害作用。MH     

Exogenous Abscisic Acid Application During Grain Filling in Winter Wheat Improves Cold Tolerance of Offspring's Seedlings

Low temperature seriously depresses seed germination and seedling growth in winter wheat (Triticum aestivum L.). In this study, wheat plants were sprayed with abscisic acid (ABA) and fluridone (inhibitor of ABA biosynthesis) at 19 days after anthesis (DAA) and repeated at 26 DAA. The seeds of those plants were harvested, and seed germination and offspring's seedling growth under low temperature were evaluated. The results showed that exogenous ABA application decreased seed weight and slightly reduced seed set and seed number per spike. Under low temperature, seeds from ABA‐treated plants showed reduced germination rate, germination index, growth of radicle and coleoptile, amylase activity and depressed starch degradation as compared with seeds from non‐ABA‐treated plants; however, activities of the antioxidant enzymes in both germinating seeds and seedling were enhanced from those exposed to exogenous ABA, resulting in much lowered malondialdehyde (MDA) and H₂O₂ concentrations and production rate. In addition, the maximum quantum efficiency of photosystem II was also enhanced in ABA‐treated offspring's seedlings. It is concluded that exogenous ABA treatment at later grain‐filling stage could be an effective approach to improve cold tolerance of the offspring during seed germinating and seedlings establishment in winter wheat.     

赤霉酸和缩节胺对转Bt基因抗虫棉棉铃Bt毒蛋白表达及氮代谢的影响

以泗抗1号和泗杂3号为研究材料,于盛花结铃期,应用赤霉酸(GA3)、缩节胺(DPC)涂抹幼铃,探讨其对棉铃Bt毒蛋白表达及氮代谢影响。结果表明,DPC能显著提高花后10d和30d(DPA)棉铃Bt毒蛋白含量,GA3降低10DPA铃壳和子棉中Bt毒白含量,但提高30DPA棉铃Bt毒蛋白含量。DPC提高了硝酸还原酶(NR)、谷氨酸-丙酮酸转移酶(GPT)、谷氨酸-草酰乙酸转移酶(GOT)活性并增加了游离氨基酸、可溶性蛋白及全氮含量,GA3处理则降低了10DPA铃壳、棉子中上述酶活性和物质含量。相关分析表明,棉铃中Bt毒蛋白含量与NR、GPT和GOT活性呈极显著正相关(r1=0.8776**,r2=0.7172**,r3=0.7028**)。可见,转Bt基因抗虫棉开花后应用DPC或GA3可提高充实期棉铃,能促进棉铃氮代谢水平,从而促进Bt毒蛋白的表达。     

不同小麦品种(系)的存放种子活力比较研究

为发掘利用高活力小麦种质材料,揭示不同小麦品种间(系)的种子活力差异及其内在生理原因,本研究以置于常温种子库老化6年的180个小麦品种(系)的种子为材料进行了种子活力比较研究。结果表明:不同小麦品种(系)间发芽率存在显著差异,其中35个小麦品种(系)的种子完全失活、不能发芽,种子活力最高的3个品种分别为豫麦18(92%),西农3517(76%)和浏虎98(72%);测定了种子浸出液电导率、种子含水量和籽粒品质性状并与发芽率进行了相关分析,发现发芽率与电导率和籽粒淀粉含量呈极显著负相关,与含水量呈显著负相关,但与籽粒总蛋白质含量相关不显著;进一步对种子活力高低差异显著的6个品种(系)进行了种子萌发关键激素赤霉素(GA3)和脱落酸(ABA)的测定,发现3个高活力小麦品种(系)(豫麦18、西农3517、浏虎98)种子中GA3/ABA的比值均显著高于3个低活力品种(系)(西农2003、洛麦21和陕麦107)。本研究表明,种子活力性状在小麦种质资源中存在广泛变异,植物内源激素GA3和ABA的相对含量是影响种子活力高低的关键因素。     

谷子SiPRR37基因对光温、非生物胁迫的响应特点及其有利等位变异鉴定

从谷子品种延谷11号克隆生物钟基因SiPRR37,通过生物信息学分析、组织特异性表达分析、4种不同光温组合条件的昼夜表达模式分析以及对NaCl、ABA、PEG、低温、Fe 5种非生物胁迫的响应特点分析,揭示SiPRR37参与谷子光温互作调控以及应对非生物胁迫的作用机制;并对160份谷子材料重测序检测SiPRR37基因的突变位点进行单倍型分析,探究该基因对谷子主要农艺性状的影响。结果表明,SiPRR37基因蛋白质编码区(sequence coding for amino acids in protein,CDS)全长2247 bp,编码748个氨基酸,含有REC和CCT 2个结构域,基于PRR37蛋白的系统进化分析发现,谷子与糜子、高粱、玉米亲缘关系最近;启动子预测分析发现,SiPRR37启动子区存在光、温、生长素、赤霉素、脱落酸、茉莉酸甲酯、干旱和盐胁迫等多种应答元件。SiPRR37相对表达量从高到低依次为根、穗颈、穗、顶叶、次顶叶、茎秆;4个光温组合条件SiPRR37均只在光照期出现1个表达峰,无论高温(27℃)还是低温(22℃),短日照相比长日照表达峰均要提前,无论长日照还是短日照,低温(22℃)相比高温(27℃)表达峰均要提前;NaCl、低温(15℃)胁迫能够抑制SiPRR37表达,PEG模拟干旱胁迫和Fe胁迫能够诱导SiPRR37基因表达,SiPRR37参与了ABA信号传导过程。位于SiPRR37 CDS区的10个SNP将160份谷子材料分为19个单倍型,其中3个单倍型(Hap_7、Hap_10、Hap_19)是改善穗部性状的有利单倍型。谷子SiPRR37基因表达具有昼夜节律性,同时受光周期和温度调控,并且参与了谷子对盐胁迫、低温胁迫、干旱胁迫和铁胁迫的应答反应,同时SiPRR37与抽穗期和多个穗部性状相关,在开展谷子高产分子辅助选育中具有一定应用潜力。     

水稻ABA生物合成基因OsNCED3响应干旱胁迫

9-顺-环氧类胡萝卜素双加氧酶(NCED)是植物内源脱落酸(ABA)生物合成的限速酶,由NCED基因家族编码,水稻中响应干旱胁迫并以此调节ABA水平的OsNCED基因尚见未报道。本研究发现在水稻已报道的5个OsNCED基因中,OsNCED3的表达受干旱胁迫诱导,复水处理后其表达快速下调,其表达模式与此过程中内源ABA含量变化趋势一致。OsNCED3的RNAi转基因植株表现为干旱敏感,且生物量下降;而过量表达OsNCED3基因增加了水稻的抗旱性。干旱胁迫下过量表达OsNCED3的转基因株系有较高的ABA水平,同时其抗氧化酶超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性,以及逆境响应基因脱水素蛋白(Dehydrin)和胚胎发育晚期丰富蛋白(LEA)转录表达均高于野生型。下调表达OsNCED3的转基因株系则呈现相反的变化趋势。因此,OsNCED3是水稻干旱胁迫响应基因,调节了干旱环境下ABA水平和抗逆性。     

棉花4个脂肪酸合成相关基因的克隆和表达特征分析

脂肪酸合成相关代谢在控制油的合成和抗非生物胁迫中均起着重要作用。其脂肪酸合成相关基因的时空表达水平直接影响油的含量和脂肪酸合成相关酶的活性。本研究克隆了4个脂肪酸合成相关基因,分别命名为GhKASII、GhKASIII、GhFAD和GhGPAT,其中GhKASIII、GhFAD和GhGPAT基因cDNA全长通过电子克隆和同源克隆得到。而GhKASII通过筛库和5'-RACE途径得到。组织表达分析表明, 上述4个基因在根、茎、叶及纤维发育不同时期均有表达,属于组成性表达基因。其中GhKASII、GhKASIII在25 DPA种子中表达量最高,GhGPAT在0 DPA胚珠和15 DPA纤维中表达量很高,GhFAD在0DPA胚珠, 15 DPA种子,20 DPA纤维中表达量均很高。不同非生物胁迫的诱导表达分析表明,上述4个基因均不同程度被茉莉酸甲酯,ABA,创伤和冷害等逆境诱导表达。     

大豆几种光周期处理效应的植物激素解析

研究了大豆光照后效应、开花逆转、光周期对源库关系的调控、摘荚后光周期反应等现象中,内源激素的变化及几种外源生长物质对大豆发育的效应。结果表明,开花前对超早熟大豆品种东农36进行短日处理,明显加快开花后发育速度,降低叶片中的ABA和ZRs含量;幼苗期(VE-V2)对晚熟品种自贡冬豆进行l0 d左右的短日处理,尔后置15 h长     

矮秆基因Rht_NM9在小麦株高建成中对内源激素含量的影响

普通小麦品种"南农9918"经甲基磺酸乙酯(ethyl methanesulfonate,EMS)诱变获得一个矮秆、多蘖、长穗突变体"NM9",在该突变体中定位到一个新的矮秆突变基因Rht_NM9。内源激素在普通小麦株高建成的过程中发挥重要的调节作用,为了解析Rht_NM9致矮的生理机制,本研究以南农9918及其矮秆突变体NM9为材料,利用酶联免疫吸附分析法(enzyme-linked immunosorbent assays,ELISA)测定了不同生育期各节间内源赤霉素(GA)、生长素(IAA)、脱落酸(ABA)和玉米素核苷(ZR)的含量,分析小麦发育关键时期的内源激素含量变化与株高的关系。结果表明,在孕穗期和抽穗期,矮秆突变体NM9中GA、ABA含量均显著高于南农9918,而ZR含量则显著低于南农9918,IAA含量在南农9918和突变体NM9之间无明显差异。此外,突变体NM9各节间中GA/ABA比值显著高于南农9918,而IAA/ABA、(IAA+GA)/ABA、ZR/ABA比值显著低于南农9918。以上结果表明小麦株高受多种激素调控,突变体中内源ABA含量升高,IAA/ABA和ZR/ABA比值降低会抑制植物株高伸长。     

Abscisic acid, gibberellin and cell viability in cereal aleurone

The aleurone layer of cereals is a secretory tissue whose activity is regulated by abscisic acid (ABA) and gibberellins (GAs). Whereas GA triggers enzyme synthesis and secretion and initiates a program that culminates in cell death, ABA prevents enzyme production and cell death. Reactive oxygen species (ROS) are key players in regulating cell viability and GA sensitizes the aleurone cell to ROS. Sensitivity of GA-treated cells results in part from a reduction in steady-state amounts of mRNAs encoding enzymes that scavenge ROS. mRNAs encoding catalase, superoxide dismutase and ascorbate peroxidase are almost undetectable in aleurone layers 24 h after incubation in GA. For layers incubated in ABA, however, the amounts of these mRNAs increase. Western blotting and enzyme activity assays confirm that GA but not ABA reduced the amount and activity of ROS scavenging enzymes (Fath et al., 2001b). Substantial amounts of ROS are produced by enzymes engaged in lipid metabolism, and by the electron transport chain in the mitochondria. Aleurone layers contain abundant stores of triglycerides and ROS are produced as these lipids are rapidly converted to sugars. We hypothesize that the ROS produced in GA-treated aleurone cells bring about cell death by disrupting the plasma membrane. Aleurone cells incubated in ABA, on the other hand, are better able to maintain redox balance. ABA does not initiate rapid triglyceride metabolism, and the activities of ROS-scavenging enzymes remain high in ABA-treated cells. We conclude that GA initiates a metabolic cascade in aleurone cells that results in death from ROS. ABA maintains viability by keeping ROS under control. This revised version was published online in August 2006 with corrections to the Cover Date.     

ABA与GA对水稻籽粒灌浆的调控

灌浆初期籽粒中ABA(脱落酸)含量(ng g-1FW)和ABA与GA(赤霉素)的比值，强势粒高于弱势粒，籽粒充实度好的组合高于籽粒充实度差的组合。开花后2天，喷施外源ABA，籽粒中ABA含量和ABA/GA增大，喷施外源GA后籽粒中GA含量增加，ABA/GA减小。低浓度(15mg/L)ABA处理后，灌浆初期籽粒中ADPG焦磷酸酶和淀粉合成酶活性及淀粉含量增加，     

不同脱水剂对杂交水稻制种种子质量及基因表达的影响

种子脱水干燥是杂交水稻制种过程的一个重要环节,种子自然脱水速率慢,遇雨天还易引起成熟期推后和穗发芽等问题。利用脱水剂快速降低种子水分对种子安全生产有重要意义。本研究以Y两优689杂交水稻品种为材料,比较不同脱水剂对其制种种子脱水及种子质量的影响。11种脱水剂均能加快种子成熟后期脱水,7号和9号脱水剂喷施后5 d,种子水分分别下降4.6%和3.6%,且对种子千粒重、发芽和幼苗生长无不良影响,种子可溶性蛋白、可溶性糖、ABA和GA3含量则显著高于对照。种子室温贮藏6个月后,与对照相比,7号、8号和9号脱水剂处理对种子发芽和幼苗生长无显著影响,7号和9号脱水剂处理的种子可溶性蛋白含量、α-淀粉酶活性、ABA含量与对照无显著差异。7号比9号的脱水效果好,可作为Y两优689的脱水剂。种子贮藏前,7号、8号和9号脱水剂处理能提高种子中OsNECD1和OsNCED2基因的表达,降低OsGA2ox1的表达;而种子贮藏6个月后,OsNECD1和OsNCED2表达量下降,OsCYP707A5和OsGA2ox1表达量上升。这表明经过6个月贮藏,脱水剂处理对种子ABA合成相关基因和GA3分解相关基因表达的影响减弱,同时OsCYP707A5的高表达导致种子ABA含量减少,可能是种子贮藏后萌发力提高的主要原因。