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1.
于2020年5月—2021年2月以苹果野生型WT植株和MdGH3-2/12的RNA干扰转基因植株(RNAi)为材料,采用水培试验方法测定了盐胁迫条件下苹果植株的表型、生长量、光合参数、叶绿素含量、抗氧化酶及活性氧(ROS)含量变化,初步解析了MdGH3-2/12在苹果响应盐胁迫中的功能。研究结果表明:(1)与WT植株相比,RNAi植株叶表面出现更多的褐斑和坏死,且根系活力显著降低,其3个株系(GR-1,GR-9,GR-10)根系活力分别为WT植株的93.8%、77.5%和90.0%;植株的生长和光合作用受到更大影响,Pn、Ci和Gs都显著降低,其中Pn值分别下降至WT植株的62.9%、77.41%和83.6%。最大光化学效率(Fv/Fm)也显著下降,3个株系分别为WT植株的97.0%、96.4%和97.5%;叶绿素含量分别显著下降至WT植株的90.7%、86.2%和81.9%,MDA含量分别为WT植株的1.15、1.25倍和1.16倍;REL含量分别为WT植株的1.23、1.39倍和1.48倍;叶片气孔开度在盐胁迫下也分别缩小了35.0%、39.0%、55.0%和46.2%。(2)与WT植株相比,RNAi植株株系体内Pro含量显著增加,3个株系分别为WT植株的7.1、10.3倍和6.3倍;叶片中ROS积累显著高于WT植株,H2O2含量分别为WT植株的1.6、1.8倍和1.8倍,而抗氧化酶活性显著下降,各材料(WT,GR-1,GR-9,GR-10)盐胁迫组的SOD酶活性分别为对照组的2.4、1.2、1.7倍和1.9倍,这表明与WT植株相比,盐胁迫下RNAi植株各株系不能有效清除ROS,对植株造成了更大伤害。(3)盐胁迫后各材料植株的Na+/K+比值分别为0.497、0.558、0.525和0.577,RNAi植株株系显著高于WT植株,但是相关离子转运基因的表达量显著低于WT植株,致使Na+不能及时外排,植株受到钠离子毒害,从而导致其耐盐性降低。以上结果表明,MdGH3-2/12在苹果植株应对盐胁迫方面发挥着重要的作用,将MdGH3-2/12[STBZ]基因干扰后苹果植株在盐胁迫下受到的伤害更大,植株对盐胁迫的抗性下降。  相似文献   

2.
为明确二肽基肽酶V(dipeptidyl-peptidase V,DppV)在稻瘟菌Magnaporthe oryzae中的生物学功能,采用在线软件对稻瘟菌DppV进行生物信息学分析,利用实时荧光定量PCR(quantitativereal-time PCR,qRT-PCR)技术分析其在稻瘟菌分生孢子不同萌发时间的表达量,通过基因敲除和功能回补、致病力测定等方法对其功能进行分析。结果表明,稻瘟菌DppV蛋白含有信号肽,定位于胞外,不含跨膜结构域,没有GPI锚定位点,为经典分泌蛋白,含DAP2结构域;其与小麦顶囊壳Gaeumannomyces tritici、早熟禾巨座壳Magnaporthiopsis poae、新烟曲霉Aspergillus novofumigatus和巴斯德伊蒙菌Emergomyces pasteurianus蛋白序列的相似度较高,分别为68.92%、68.41%、51.09%和49.96%;在稻瘟菌分生孢子萌发早期DppV基因均有表达,但当萌发时间为24 h时,其相对表达量最高,为5.61;稻瘟菌DppV基因敲除突变体菌株的菌落生长速度明显下降,产孢量(8.18×104个/cm2)和4个产孢相关基因MoHox2、MoCon8、MoSec22MoCos1的相对表达量显著降低,分生孢子萌发率(56.21%)明显下降,对SDS、NaCl、山梨醇和H2O2胁迫更加敏感,对水稻的致病力显著降低,水稻植株内真菌生物量明显减少,而稻瘟菌DppV基因回补菌株的表型、耐胁迫能力和致病力等则恢复到稻瘟菌野生型菌株的水平。表明DppV基因在稻瘟菌营养生长、产孢、致病力、对外界胁迫的应答及其在水稻体内的定殖等方面起着重要作用。  相似文献   

3.
为明确球孢白僵菌Beauveria bassiana定殖对番茄植株抗病性的影响及作用机理,采用灌根法将球孢白僵菌芽生孢子悬浮液接种于番茄植株内,并通过人工接种灰霉菌Botrytis cinerea评价球孢白僵菌定殖后番茄对灰霉病的抗性水平;检测灰霉菌胁迫下番茄植株不同位置叶片内球孢白僵菌的相对含量;测定番茄叶片内草酸氧化酶(oxalate oxidase,OXO)、几丁质酶(chitinase,CHI)和ATP合成酶(ATP synthase,atpA)3种抗病相关基因的表达量。结果表明,球孢白僵菌定殖能够提高番茄植株对灰霉病的抗性,接种灰霉菌第5天,番茄植株发病率、病斑直径和病情指数分别下降了61.6%、41.4%和26.4%;在灰霉菌胁迫下番茄植株内球孢白僵菌偏好于在病原菌感染位置定向聚集,并且引起植物抗病基因OXOCHIatpA的表达量上调。表明球孢白僵菌能通过内生定殖与植物互作提高植物抗病性,在植物病害生物防治领域有较大应用潜力。  相似文献   

4.
为明确南方根结线虫Meloidogyne incognita效应蛋白MiV901在其寄生过程中的生物学功能,通过构建MiV901基因的植物表达载体,利用根癌农杆菌Agrobacterium tumefaciens介导的花序浸染法将其转化拟南芥Arabidopsis thaliana,并采用室内人工接种法测定转基因植株对灰葡萄孢 Botrytis cinerea侵染及南方根结线虫寄生的影响。结果显示:经Southern blot检测,MiV901基因以不同的拷贝数插入到转基因拟南芥株系901-6、901-8和901-12的基因组中,且qPCR检测结果证实 MiV901基因能够正常表达。3个转基因拟南芥株系901-6、901-8和901-12叶部接种灰葡萄孢3 d后,叶片上形成的病斑平均直径分别为1.00、1.06、1.05 cm,比野生型对照扩大了9.9%~16.5%。相比野生型对照,转基因拟南芥株系901-12、901-6和901-8接种南方根结线虫2龄幼虫后根系上产生了更多的雌虫和卵块,雌虫数分别显著增加了45.4%、34.4%和23.7%,卵块数分别显著增加了51.2%、 46.3%和31.7%。表明异源表达MiV901基因能够抑制植物免疫,增加拟南芥对灰葡萄孢和南方根结线虫侵染的敏感性。  相似文献   

5.
为了研究马铃薯EPFL9/StSTOMAGEN基因在气孔发育过程中的功能,从马铃薯品种大西洋中克隆得到气孔密度表皮模式因子StSTOMAGEN,将其通过农杆菌介导的遗传转化方法,在拟南芥中异位表达,对该基因的表达特性及功能进行分析。结果显示:StSTOMAGEN主要定位于细胞间隙和细胞核,并且它在顶端未展开叶中表达最多,表达量在1 d中呈现周期性变化,长日照条件下(16 h/8 h,光/暗),在授时因子为20 h时表达量最高;聚乙二醇(PEG)模拟干旱胁迫处理4 h后其表达量达到最高,而脱落酸(ABA)和NaCl胁迫处理后,其表达水平呈现出持续降低的趋势。功能鉴定表明,过表达StSTOMAGEN基因的拟南芥(ST)相对于野生型(WT)气孔密度增大63%~83%,光合速率增大36%~42%。此外,ST植株离体叶片的水分散失速率增大24%~55%,离体叶片H2O2含量也显著升高,约为野生型的50%~100%。干旱胁迫下,ST株系的光合速率、Fv/Fm和瞬时水分利用效率都显著低于野生型,其中光合速率降低55%~66%,Fv/Fm降低22%~50%,瞬时水分利用效率降低11%~12%。综上结果可知,表皮模式因子StSTOMAGEN通过正调控气孔密度而降低植物的抗旱性,该结果为后期通过基因编辑技术降低StSTOMAGEN基因的表达,培育抗旱节水型马铃薯新品种奠定理论基础。  相似文献   

6.
我国南方稻区水稻骨干亲本纹枯病抗性鉴定与分析   总被引:3,自引:0,他引:3  
运用改进的纹枯病菌接种法,对当前我国南方稻区17个骨干亲本纹枯病抗性进行了田间鉴定,测定和分析了各材料的纹枯病病级(RD)、相对病斑高度(HR)及株高(HP)。通过试验,将17个骨干亲本分为抗病、中等抗病、中等感病和感病4种类型;筛选出了3个抗病亲本(明恢63、蜀恢527、广恢128)和3个中抗亲本(绵恢725、珍汕97B和协青早B);供试亲本的平均RD为4.78,平均HR为0.52;通过分析,发现纹枯病病级(RD)和相对病斑高度(HR)呈极显著正相关(y=8.373x+0.435,y=RD;x=HR)、株高和病级相关不显著。  相似文献   

7.
蜡蚧轮枝菌生物学特性及其对温室白粉虱致病性的研究   总被引:5,自引:2,他引:5  
通过对引进的 6个蜡蚧轮枝菌的生长、产孢量、分生孢子萌发和对温室白粉虱若虫侵染致病性的研究表明 ,6个菌株在 10~30℃下均可生长 ,菌丝生长和产孢的最适温度为 25℃ ,但菌株间存在显著差异。菌株V4和V5产孢量显著高于其他菌株 ,14d的产孢量分别为 2.6×109个 /皿和 2.3×109个 /皿。菌株V4V1的菌落直径累计增长量和日增长量显著地高于其他菌株 ,分别为43.7、2.85mm和43.1、2.84mm。温度对孢子的萌发影响较大 ,15℃时孢子萌发缓慢 ,萌发达 90%以上时需 24h ,20~30℃下菌株V2 孢子萌发速率明显高于其他菌株 ,10h萌发率达 90%以上 ,30℃下 ,菌株V6萌发速度明显延缓。对温室白粉虱若虫的致病性 ,菌株间存在一定的差异。V2V4接菌后第 5天若虫死亡率分别为 70.2%和 63.5% ,综合其菌丝生长、产孢量、萌发率和致病性表明 ,菌株V2V4可作为生物制剂的生产菌株  相似文献   

8.
SA诱导拟南芥对灰霉病的抗性与木质素含量的关系   总被引:1,自引:0,他引:1       下载免费PDF全文
对拟南芥植株喷施水杨酸后,接种灰霉病菌(Botrytis cinerea),观察植物对病原物反应的变化。结果表明:喷施水杨酸可以降低拟南芥灰霉病的发生率。在平板培养中,水杨酸对灰霉菌分生孢子的萌发及菌丝的生长均无明显抑制作用,因此认为水杨酸处理后发病率的下降是由于水杨酸处理提高了植物的抗病性,且诱导抗性的持久期在10 d以上。观察中还发现在拟南芥叶片上喷施SA或接种灰霉菌都会使叶片PAL和POD活性上升,进而引起木质素的积累,两者在积累量上和时间程度上具较好的一致性,说明水杨酸诱导拟南芥抗灰霉病的过程中,通过PAL和POD活性的提高进而引起木质素含量的增加,可能是诱导拟南芥产生抗性的机制之一。  相似文献   

9.
以‘东方不败’、‘常绿’和‘C069’3种萱草(Hemerocallis sp.)为材料,研究了不同浓度的外源脱落酸(ABA)和过氧化氢(H2O2)对植物抗旱性的影响。结果表明,ABA和H2O2对3个萱草品种的抗旱性呈现明显的剂量效应,当浓度适宜时能提高其抗旱性。干旱条件下喷施200 μmol· L-1ABA,‘东方不败’和‘C069’比其它处理保持了更高的叶片RWC和气孔密度以及更低的叶片相对电导率和气孔开度。可见,200μmol·L-1 ABA和H2O2对提高‘东方不败’和‘C069’抗旱性的效果最为明显;而500 μmol·L-1 ABA和H2O2处理显著地提高‘常绿’的抗旱性.初步推断,ABA和H2O2均可能包含在萱草抗旱的信号途径中,但是两者的信号功能及其关系有待进一步研究。  相似文献   

10.
为明确蚜虫内共生细菌Buchnera能否协助蚜虫抑制植物的防御反应,采用混合抗生素除去瓜蚜Aphis gossypii体内的Buchnera,采用刺吸电位技术和生命表方法比较除菌瓜蚜和正常瓜蚜在黄瓜上的适合度,并测定被除菌瓜蚜和正常瓜蚜预侵染黄瓜叶片中H2O2、茉莉酸和水杨酸的含量变化以及抗蚜特性变化。结果表明,除菌瓜蚜体重较正常瓜蚜极显著下降了71.15%,净增殖率极显著下降了95.60%,但寿命延长了2.34倍;刺吸电位图谱显示除菌瓜蚜几乎不产生代表唾液分泌的E1波和韧皮部被动吸食的E2波。在黄瓜叶片中,除菌瓜蚜比正常瓜蚜能激发产生更高水平的H2O2;在为期14 d的侵染中,除菌瓜蚜侵染诱发总茉莉酸含量持续升高、总水杨酸含量持续下降,而正常瓜蚜侵染诱发总茉莉酸含量先升高后降低、总水杨酸含量先降低后升高。在被除菌瓜蚜预侵染黄瓜上,瓜蚜进入稳定取食的时间显著滞后于正常瓜蚜预侵染处理,且前3 d的产蚜量显著低于正常瓜蚜预侵染处理。表明Buchnera除了可为蚜虫提供必需的营养外,还有调控和抑制寄主植物防御反应的功能。  相似文献   

11.
为筛选出橡胶树白粉菌Oidium heveae Steimnann新的寄主载体,通过接种野生型拟南芥Col-0、突变体Sr1-4D及eds1,观察了橡胶树白粉菌侵染不同拟南芥突变体的过程,PCR扩增测序法验证相关致病基因,构建橡胶树白粉菌-拟南芥互作体系。结果表明,在Col-0、Sr1-4D叶片上面,橡胶树白粉菌产生部分菌丝后停止生长,不能形成典型的橡胶树白粉病症状;但能成功侵染eds1叶片,在叶片的正、背面有银白色辐射状菌丝,后期在病斑上出现一层粉层,表现橡胶树白粉病的典型症状。组织染色和显微观察结果显示,在突变体eds1叶片上橡胶树白粉菌完成了侵染过程。PCR扩增测序结果表明接种后突变体eds1叶片上及组织中病菌均为橡胶树白粉菌;使用橡胶树白粉菌3个致病相关基因作为靶标,验证了eds1和橡胶树上白粉菌基因组中的3个致病相关基因相似度均达到99%~100%。表明橡胶树白粉菌可侵染拟南芥突变体eds1。  相似文献   

12.
In Arabidopsis, abscisic acid (ABA) application can induce resistance by priming for callose deposition; this resistance is impaired in ABA-deficient and -insensitive mutants. In tomato, ABA-deficiency causes resistance to Botrytis cinerea. Here, we show that callose deposition after B. cinerea inoculation is weaker in the ABA-deficient sitiens tomato mutant compared to the wild type (WT). Inhibition of callose synthesis did not affect resistance in sitiens, but caused additional susceptibility in WT. These findings indicate that callose deposition is not part of sitiens defence responses that are effective in blocking B. cinerea and suggest that callose deposition only contributes to WT basal resistance. Furthermore, also in tomato callose formation is at least in part ABA-dependent. However, it seems that in contrast to Arabidopsis, basal ABA levels in tomato are sufficiently high to prime for callose deposition.  相似文献   

13.
Systemic acquired resistance (SAR) is induced following inoculation of Peronospora tabacina sporangia into the stems of Nicotiana tabacum plants highly susceptible to the pathogen. Previous results have shown that accumulation of acidic β-1,3-glucanases (PR-2's) following induction of SAR by P. tabacina may contribute to resistance to P. tabacina. We showed that up-regulation of the PR-2 gene, PR-2d, following stem inoculation with P. tabacina, is associated with SAR. Studies using plants transformed with GUS constructs containing the full length promoter from PR-2d or promoter deletions, provided evidence that a previously characterized regulatory element that is involved in response to salicylic acid (SA), may be involved in regulation of PR-2d following induction of SAR with P. tabacina. This work provides evidence that regulation of PR-2 genes during P. tabacina-induced SAR may be similar to regulation of these genes during infection of N-gene tobacco by TMV or following exogenous application of SA, and provides further support for the role of SA in regulation of genes during P. tabacina-induced SAR.  相似文献   

14.
The Arabidopsis MPK3 gene product participates in disease resistance mediated by the MAP kinase cascade. The expression of the MPK3 gene is induced by pathogen inoculation and treatment with chemicals such as salicylic acid (SA) and methyl jasmonate (JA), but the detailed expression pattern of the MPK3 gene has been largely unknown. To investigate MPK3 gene expression in response to disease stress, we fused the MPK3 promoter to the firefly luciferase gene to create a real-time monitoring system for regulated gene expression in planta. The results of an in vivo reporter assay using transgenic Arabidopsis plants harboring MPK3::Fluc showed that the MPK3 promoter activity was induced by treatment with chemicals such as SA and benzo(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester (BTH), that induce defense gene expression. Inoculation with the fungal pathogen Botrytis cinerea resulted in systemic induction of MPK3::Fluc.  相似文献   

15.
史佳琪  强胜  张裕 《植物保护学报》2023,50(5):1347-1357
为提升齐整小核菌Sclerotium rolfsii菌株SC64作为生物除草剂的应用潜力,解析植物对其的防御机制,通过比较6种生态型拟南芥Arabidopsis thaliana(Col-0、Ms-0、Gr-1、Se-0、Tu-0和Wa-1)被菌株SC64侵染后病理特征、茎秆木质素水平以及抗病相关基因表达的差异,筛选出抗性最高和抗性最弱的典型生态型拟南芥,通过施加0.1 mmol/L外源水杨酸来验证其在拟南芥防御菌株SC64侵染过程中对木质素代谢及病理的调节作用,并比较分析6种生态型拟南芥水杨酸上游调控通路基因MPK4启动子区域的甲基化差异。结果显示,6种生态拟南芥中Col-0的木质素化程度最高,被菌株SC64侵染时木质素代谢通路基因CAD4和PAL3表达上调最显著,与其抗病能力最强相吻合,而Ms-0的病理表型和抗病能力最弱。外源水杨酸预处理使具典型抗性的生态型拟南芥Col-0受菌株SC64侵染后发病程度加重,木质素化程度减弱,同时木质素代谢通路基因MYB46、LAC17、PAL3和CAD4被菌株SC64侵染后的表达上调程度均显著减弱。另外,6种生态型拟南芥MPK4基因启动子区域CHH...  相似文献   

16.
Tomato and transgenic oilseed rape plants expressing the Cf-9 resistance gene develop a hypersensitive response (HR) after injection of the corresponding Avr9 gene product. It was investigated whether induction of a HR conferred resistance to different fungal pathogens in tomato and oilseed rape. Induction of an AVR9 mediated HR at the pathogen infection site delayed the development of the biotrophs Oidium lycopersicum in tomato and Erysiphe polygoni in oilseed rape, but enhanced the development of the necrotrophs Botrytis cinerea and Alternaria solani in tomato and Sclerotinia sclerotiorum in oilseed rape. Interestingly, delayed fungal disease development was observed in plant tissues surrounding the HR lesion regardless of whether a necrotrophic or biotrophic pathogen was used. In tomato, AVR9 injection induced systemic expression of PR1, PR2 and PR3 defence genes but did not induce systemic resistance to O. lycopersicum, B. cinerea or A. solani. In oilseed rape, AVR9 injection temporarily induced systemic resistance to Leptosphaeria maculans and E. polygoni, but did not induce detectable systemic expression of PR1, PR2 or Cxc750. These results give new insights into the potential uses of an induced HR to engineer disease resistance.  相似文献   

17.
为明确广谱性抗病毒基因—酵母pac1基因对葡萄B病毒(Grapevine virus B,GVB)的抗性效果,通过农杆菌介导的遗传转化,将pac1基因导入西方烟37B,对转基因植株进行PCR鉴定及Southern blot分析,通过病毒摩擦接种观察症状以及实时荧光定量RT-PCR检测植株体内病毒含量,并对转基因植株抗病性进行初步鉴定。结果表明,目的基因pac1成功导入并整合至西方烟37B基因组,共获得10个转基因株系。不同株系的T1代烟草中阳性植株比例为16.7%~72.7%,表明目的基因可成功遗传到子代。接种病毒后转基因植株普遍延迟发病,但后期症状与非转基因对照相似,其中仅1个转基因株系B6具有不表现典型症状等抗性反应。接种植株病毒含量检测中,所有转基因植株均检测到病毒存在,但表现为抗病的B6株系中病毒含量显著低于非转基因对照,表明该转基因植株虽不能完全抵抗GVB侵染,但对GVB具有耐病性。  相似文献   

18.
Flax engineering to yield increased resistance to pathogens is the goal of this study. Since carotenoids act as antioxidants it is thus postulated that the accumulation of a higher quantity of these compounds in the transgenic plants might improve their resistance to pathogen infection.Our approach was based on the generation of transgenic flax overproducing carotene and analysis of its susceptibility to Fusarium infection. For transformation bacterial gene – crtB was used. As expected, transgenic plants showed increased resistance against pathogen infection.The impact of carotenoids on plant resistance to infection was verified by generation and analysis of transgenic flax with decreased content of carotene. The transgenic plants were obtained by suppression of endogenous flax gene coding for lycopene β-cyclase. Plant analysis revealed decrease in carotene content, however, an unexpected increase in resistance against Fusarium infection was detected. Further analysis of metabolites in the plants revealed that an increase in accumulation of other terpenoids and tocopherols, squalene and menthol were among them. Thus, it is suggested that repression of carotene synthesis results in the redirecting of substrates to other branches of isoprenoids synthesis.We conclude that a general level of antioxidants rather than the presence of any particular compound is the most important factor in resistance of the flax plant to pathogen infection.  相似文献   

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