超表达OsPR1A增强了Xa21介导的水稻对白叶枯病的抗性反应     

刘玉晴  燕高伟  张彤  兰金苹  郭亚璐  李莉云  刘国振  窦世娟 《中国农业科学》2021,54(23):4933-4942

【背景】前期研究发现,水稻病程相关蛋白质OsPR1A的表达受上游抗病基因Xa21调控,接菌后早期启动Xa21介导的OsPR1A较高水平表达对水稻抵抗白叶枯病菌至关重要。同时OsPR1A也受到水稻白叶枯病菌（Xanthomonas oryzae pv. oryzae,Xoo）的诱导表达。对于OsPR1A的研究绝大部分是作为抗性反应发生的标志基因佐证其他基因或途径在抗性中的作用,缺乏直接的证据证实OsPR1A本身的生物学功能。【目的】通过获得OsPR1a-OX超表达转基因植株,调查其表型及农艺性状,并明确OsPR1A蛋白质表达与抗性的关系,为鉴定OsPR1A功能提供依据。【方法】通过农杆菌介导法,将构建的OsPR1a-OX转化载体转入到水稻受体4021中,利用PCR和免疫印迹（western blot,WB）技术分别在基因水平和蛋白质水平上筛选并鉴定OsPR1A超表达阳性纯合株系。在成熟期,调查OsPR1A超表达转基因植株的表型及农艺性状（株高、穗长、分蘖数、结实率和籽粒大小等）。在31℃条件下,将生长2周的水稻幼苗TP309、4021和OsPR1A超表达转基因植株接种水稻白叶枯病菌,并在接菌0、2、4、6、8、10和12 d时测量病斑长度。在接菌0、4和6 d时,收集TP309、4021和OsPR1A超表达转基因植株的水稻叶片,提取蛋白质,利用WB技术检测OsPR1A的表达特征。【结果】构建了OsPR1a-OX转化载体,并转入到受体4021中,筛选并鉴定到2个OsPR1A超表达转基因纯合株系（#704和#709）。调查了OsPR1A超表达转基因植株在成熟期的表型及农艺性状,与对照4021相比,#704和#709的株高较矮、穗长较短、分蘖数减少、结实率降低,但籽粒稍大,可能与结实率低有关。在31℃条件下,OsPR1A超表达转基因植株的病斑长度与对照4021相比明显缩短,结果具有显著性差异（P<0.05）。在接菌0、4和6 d的材料中,超表达转基因植株#704和#709中OsPR1A始终有较高水平的表达丰度,从而提高了对白叶枯病菌的抗性。【结论】采用农杆菌介导法,获得OsPR1A超表达转基因植株;超表达OsPR1A影响到水稻的正常发育过程;超表达OsPR1A后增强了Xa21介导的水稻对白叶枯病的抗性。  相似文献   

中国东北地区水稻纹枯病病原菌种类及融合群的分子鉴定     

张照茹  魏松红  杨晓贺  王海宁  许月 《植物保护》2019,45(6):283-287

为明确中国东北地区水稻纹枯病病原菌种类及融合群的归属情况, 2015－2017年从黑龙江省、吉林省和辽宁省的17个水稻主产区采集水稻纹枯病标样, 分离获得水稻纹枯病菌214株, 运用水稻纹枯病菌的不同病原菌及融合群的特异性引物对214株水稻纹枯病菌进行病原菌种类和融合群鉴定, 并利用rDNA内转录间隔区(ITS)序列, 对供试水稻丝核菌的融合群归属进行了分析。结果表明:供试214株水稻纹枯病菌分属于茄丝核菌Rhizoctonia solani和水稻丝核菌Rhizoctonia oryzae-sativae, 菌株数分别为198株和16株, 占比分别为92.52%和7.48%。茄丝核菌菌株分属于2个融合群, 分别为AG1-IA和AG4, 菌株数分别为191株和7株, 占比分别为96.46%和3.54%。水稻丝核菌菌株均属于AG-Bb融合群, 菌株数为16株。不同年份水稻纹枯病的病原菌种类及融合群出现的频率和地域分布无明显变化, 而不同地域间水稻纹枯病病原菌的种类及融合群具有明显的分化特征, AG1-IA融合群在中国东北三省各个水稻产区均有分布且均为优势融合群, AG4融合群在辽宁省盘锦市出现频率最高, 水稻丝核菌AG-Bb融合群在吉林省吉林市、通化市和梅河口市出现频率最高。  相似文献   

Ray blight of pyrethrum in Australia: a review of the current status and future opportunities     

M. A. H. B. Bhuiyan  N. Vaghefi  P. W. J. Taylor 《Plant pathology》2019,68(4):620-627

Ray blight caused by Stagonosporopsis tanaceti is one of the most important diseases of pyrethrum (Tanacetum cinerariifolium), a perennial herbaceous plant cultivated for the extraction of insecticidal pyrethrins in Australia. The disease is responsible for complete yield loss in severe outbreaks. Infected seed is considered as the principal source of S. tanaceti. Infection hyphae remain only in the seed coat and not in the embryo, resulting in pre- and post-emergence death of seedlings and latent infection. Therefore, quantification of the level of infection by S. tanaceti within seed using a qPCR assay is important for efficient management of the disease. Stagonosporopsis tanaceti completes its life cycle within 12 days after leaf infection through production of pycnidia and can infect every tissue of the pyrethrum plant except the vascular and root tissues. Ray blight epidemics occur in pyrethrum fields through splash dispersal of pycnidiospores between adjacent plants. Besides steam sterilization, thiabendazole/thiram and fludioxonil are effective seed-treating chemicals in controlling S. tanaceti before planting begins. Ray blight is currently managed in the field through the foliar application of strobilurin fungicides in the first 1–2 years of crop establishment. Later on, difenoconazole and multisite specific fungicides in the next 2–3 years during early spring successfully reduce ray blight infestation. Avoiding development of resistance to fungicides will require more sustainable management of ray blight including the development and deployment of resistant cultivars.  相似文献   

马铃薯AP2/ERF转录因子的克隆及植物表达载体构建     

王慧  刘洪  杨奕琦  许思思  熊兴耀  周倩 《分子植物育种》2020,(8):2563-2568

AP2/ERF基因家族转录因子普遍存在于植物中,参与植物体内的各种生物学过程,包括植物的生长发育、生物和非生物胁迫响应等。前期转录组测序的结果发现马铃薯‘加湘1号’一个AP2/ERF家族基因(PGSC0003DMG400012154)在接种晚疫病菌(Phytophthora infestans)24 h后被显著激活。从接种P.infestans 24 h的‘加湘1号’的总RNA中通过RT-PCR获得了该基因CDS序列为885 bp,BLAST分析显示该基因编码一个295个氨基酸残基的蛋白,并且含有1个AP2/ERF结构域,是AP2/ERF转录因子家族中的ERF亚家族的一员。本研究将该基因与XcmⅠ酶切的表达载体pCXSN连接,转化大肠杆菌,通过测序挑选插入正确克隆酶切验证,并成功转化农杆菌。本研究结果为进一步研究该基因的功能提供了帮助。  相似文献   

Comparison of outcomes following treatment of septic calcaneal bursitis by needle or bursoscopic lavage: A retrospective study of 29 horses     

M. J. S. Duggan  T. S. Mair 《Equine Veterinary Education》2021,33(3):135-142

Sepsis of the calcaneal bursae (CB) presents significant treatment challenges with limited clinical data available in the literature. The objective of this retrospective cohort study was to assess the clinical outcomes associated with CB lavage using either a through-and-through needle or bursoscopic technique. Clinical records of 29 horses treated for septic calcaneal bursitis using either technique between 2005 and 2019 were reviewed. Fisher’s exact test was used to assess statistical significance between first surgical technique and success at first surgery (i.e. not requiring >1 lavage), survival to discharge and return to work (RTW). Bursoscopy was performed in 13/29 (44.8%) cases, and needle lavage in 16/29 (55.2%). In the needle group, 12 (75%) horses were discharged following the first surgery. Four had repeat interventions; two (12.5%) had needle lavage and two (12.5%) had bursoscopy. Of the two horses to have repeat needle lavage, one was subjected to euthanasia and one discharged, and of the two that underwent bursoscopy, one was discharged and one received a third bursoscopy prior to discharge. In the bursoscopy group, seven (53.8%) were discharged and three (23.1%) were subjected to euthanasia following the first surgery. Three (23.1%) received a second bursoscopic lavage with one discharged, one subjected to euthanasia and one having a third bursoscopic lavage prior to discharge. Overall, 18/24 (75%) followed up cases RTW, 10 (55.5%) from the needle group, eight (44.4%) the bursoscopy group. No statistically significant differences between first surgical technique used and success at first surgery (no subsequent lavage(s) required), survival to discharge or return to work were detected. The main limitations of this study are that it is a retrospective study, has a small population with limited statistical power and potential selection bias. No statistically significant differences existed between the outcomes of the two techniques, contrary to the belief that bursoscopic lavage is superior. Larger, multicentred studies, with greater statistical power are required to further assess this relationship.  相似文献   

Impact of weather variables and season on sporulation of Phytophthora pluvialis and Phytophthora kernoviae     

Stuart Fraser  Mireia Gomez‐Gallego  Judy Gardner  Lindsay S. Bulman  Sandra Denman  Nari M. Williams 《Forest Pathology》2020,50(2)

Phytophthora pluvialis and Phytophthora kernoviae are the causal agents of important needle diseases on Pinus radiata in New Zealand. Little is known about the epidemiology of the diseases, making the development of control strategies challenging. To investigate the seasonality and climatic drivers of sporulation, inoculum traps, consisting of pine fascicles floating on water in plastic containers, were exchanged fortnightly at five sites in P. radiata plantations between February 2012 and December 2014. Sections of needle baits were plated onto selective media and growth of Phytophthora pluvialis and P. kernoviae recorded. To explore the generalizability of these data, they were compared to detection data for both pathogens from the New Zealand Forest Health Database (NZFHDB). Further, equivalent analyses on infection of Rhododendron ponticum by P. kernoviae in Cornwall, UK allowed the comparison of the epidemiology of P. kernoviae across different host systems and environments. In New Zealand, inoculum of P. pluvialis and P. kernoviae was detected between January–December and March–November, respectively. Inoculum of both species peaked in abundance in late winter. The probability of detecting P. pluvialis and P. kernoviae was greater at lower temperatures, while the probability of detecting P. pluvialis also increased during periods of wet weather. Similar patterns were observed in NZFHDB data. However, the seasonal pattern of infection by P. kernoviae in the UK was the opposite of that seen for sporulation in New Zealand. Phytophthora kernoviae was likely limited by warmer and drier summers in New Zealand, but by colder winter weather in the UK. These results emphasize the importance of considering both environmental drivers and thresholds in improving our understanding of pathogen epidemiology.  相似文献   

基于形态学及多基因位点的枸杞褐斑病病原鉴定     

王艳  晋玲  申培增  崔治家  朱田田 《植物保护》2020,46(5):122-127

枸杞Lycium barbarum Mill.是一种药食两用的名贵中药材,褐斑病是2010年在甘肃省枸杞种植区发生的由无性态类真菌新种枸杞小黑梨孢Stigmella lycii引起的新病害,据2015年-2018年调查,该病害已扩展至甘肃省各枸杞种植区,常年发病率45%～65%,严重度2～3级。本研究首次对枸杞小黑梨孢有性态形态进行描述,并结合分子生物学方法,确定该病病原菌的系统发育地位,为该病的防治提供一定的理论依据。枸杞褐斑病菌在离体培养条件下和越冬病叶上均可形成有性态。子囊壳球形至亚球形,大小185.6μm×176.6μm,有喙,喙的大小为(35.7～53.6)μm×(32.0～53.55)μm;子囊袋状,大小(103.2～165.7)μm×(15.7～22.4)μm,含8个子囊孢子;子囊孢子砖格状,大小(25.9～36.5)μm×(9.4～14.1)μm,平均31.2μm×12.3μm,具(4～)6～7个纵隔和1～3个横隔。通过ITS、LSU、RPB2和EF1-α多基因位点联合构建系统发育树,确定该菌为子囊菌门格孢腔菌目格孢腔菌科真菌。秋末冬初清洁田园,减少来年初侵染源,是有效防治枸杞褐斑病的关键措施。  相似文献   

亚硝基谷胱甘肽还原酶 GSNOR1正调控植物对疫霉菌的抗性     

刘杏芍  宋斯雨  高贤贤  单卫星  强晓玉 《西北农业学报》2020,30(10):1457-1471

疫霉属（ Phytophthora）卵菌引致马铃薯晚疫病等作物灾难性病害，严重威胁作物的可持续生产。由于病菌毒性变异，导致品种抗病性丧失问题突出。因此，挖掘植物广谱和持久抗病基因，并探索其在抗病育种中的有效利用具有重要的科学意义。亚硝基谷胱甘肽还原酶1（GSNOR1）是植物氮信号通路中高度保守的关键还原酶，其参与调节r基因介导的植物抗性和非寄主抗性。然而， GSNOR1参与抗疫霉菌的免疫功能和作用机理尚不清楚。本研究中，借助拟南芥与寄生疫霉菌互作的模式体系，首先发现拟南芥T-DNA插入突变体 gsnor1-3对寄生疫霉菌呈现感病表型。进一步利用病毒诱导的基因沉默（VIGS）降低烟草叶片中 GSNOR1同源基因的表达量，在此基础上，通过抗病表型分析以及一系列抗性相关功能的检测，结果表明，沉默本氏烟草 GSNOR1同源基因能够在寄生疫霉菌侵染植物的过程中削弱植物体内的活性氧（ROS）迸发、病程相关基因（PR genes）的诱导表达以及MAPK信号转导，从而增强了植物对疫霉菌的感病性。本研究揭示了植物 GSNOR1对疫霉菌具有高度保守的抗性功能，并初步解析了 GSNOR1正调控植物抗疫霉菌的作用机理，为进一步探索 GSNOR1在马铃薯抗晚疫病中的功能及其在抗病育种中的有效利用奠定了重要的理论 基础。  相似文献   

Whitened kernel surface: A fast and reliable method for assessing Fusarium severity on cereal grains by digital picture analysis     

Marine Ollier  Vincent Talle  Anne‐Laure Brisset  Zo Le Bihan  Simon Duerr  Marc Lemmens  Ellen Goudemand  Olivier Robert  Jean‐Louis Hilbert  Hermann Buerstmayr 《Plant Breeding》2019,138(1):69-81

Fusarium head blight (FHB) is a cereal disease of major importance responsible for yield losses and mycotoxin contaminations in grains. Here, we introduce a new measurement approach to quantify FHB severity on grains based on the evaluation of the whitened kernel surface (WKS) using digital image analysis. The applicability of WKS was assessed on two bread wheat and one triticale grain sample sets (265 samples). Pearson correlation coefficients between Fusarium‐damaged kernels (FDK) and WKS range from r = 0.77 to r = 0.81 and from r = 0.61 to r = 0.86 for the correlation between deoxynivalenol (DON) content and WKS. This new scoring method facilitates fast and reliable assessment of the resistance to kernel infection and shows significant correlation with mycotoxin content. WKS can be automated and does not suffer from the “human factor” inherent to visual scorings. As a low‐cost and fast approach, this method appears particularly attractive for breeding and genetic analysis of FHB resistance where typically large numbers of experimental lines need to be evaluated, and for which WKS is suggested as an alternative to visual FDK scorings.  相似文献   

樟子松枯梢病的防治研究   总被引：6，自引：0，他引：6  

徐贵军  宋晓东  刘桂荣  陈江燕  吴锈钢  李淑华  吴景云 《中国森林病虫》2001,20(3):16-19

樟子松移植苗枯梢病Sphaeropsis sapinea可以通过药剂有效地进行防治。移植前以50％多菌灵wp500倍液浸苗；在人工林中，内吸性杀菌剂－甲基托布津、多菌灵为首选药剂；在树木新梢生长停止期和展叶中期（辽西北为6月初和6月中旬），70％甲基托布津wp 1000倍液或50％多菌灵wp 500倍液防治2次。保护性杀菌剂－百菌清效果不佳。适时的卫生伐对该病有明显的抑制作用。  相似文献