超表达OsPR1A增强了Xa21介导的水稻对白叶枯病的抗性反应

【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。     

Banishing barberry: The history of Berberis vulgaris prevalence and wheat stem rust incidence across Britain

Wheat stem rust, caused by the fungus Puccinia graminis f. sp. tritici (Pgt), is a notoriously damaging disease of wheat and barley. Pgt requires two hosts to complete its lifecycle; undergoing asexual reproduction on cereal crops and completing sexual reproduction on Berberis spp. The latter stage of its lifecycle is of particular importance in temperate regions such as western Europe, where asexual urediniospores are unable to survive cold winter weather. In the past, the crucial role of Berberis in the lifecycle of stem rust led to intensive eradication campaigns, initially carried out by farmers in the face of hostile scientific opinion. In the United Kingdom, common barberry (Berberis vulgaris) is today a relatively rare plant. Stem rust is, however, currently experiencing a resurgence; at the same time, there has been a general increase in the prevalence of barberry and an upsurge in its planting which, in the United Kingdom, is associated with attempts to encourage the endangered barberry carpet moth (Pareulype berberata). This article situates current developments within a broader chronological framework, examining changing attitudes towards barberry and rust in England in the past and the history of the plant's use and cultivation. It assesses how widespread B. vulgaris really was in the environment historically, and thus the scale of its eradication. We suggest that Berberis was never widely established as an archaeophyte in the United Kingdom. Current attempts to re-establish it are based on a misunderstanding of the plant's historical status and could potentially pose a serious threat to food security.     

High vegetative compatibility diversity of Cryphonectria parasitica infecting sweet chestnut (Castanea sativa) in Britain indicates multiple pathogen introductions

Chestnut blight, caused by Cryphonectria parasitica, was identified in Devon, UK, in December 2016. Intensive surveys detected the disease at further sites in Devon (seven), Berkshire (one), Dorset (one), Derbyshire (four) and a cluster of eight sites in southeast London. Over 570 survey samples were tested, and 227 were positive for C. parasitica by isolation and real-time PCR. A total of 227 isolates were tested for mating type, and 197 screened for vegetative compatibility group (VCG) and compared with VCGs known from mainland Europe. The same isolates were also screened for the presence of Cryphonectria hypovirus 1 (CHV-1). Eleven VCGs were identified within the UK population. Five corresponded to already known European VCGs but six were unique. The European VCGs mainly came from the Devon, Dorset, Berkshire and Derbyshire disease outbreaks, whilst unique VCGs were almost exclusively from the southeast London cluster. Both mating types were detected, but only one mating type was present at each site, with the exception of a single Devon site. Perithecia of C. parasitica were never observed at any site. CHV-1 was found in seven isolates from three different locations and was always subtype-I, which has limited hypovirulence. Therefore, although CHV-1 is associated with C. parasitica at some outbreaks, it probably has limited impact on virulence. The diversity of VCGs and their distribution at outbreak sites, together with findings of CHV-1, suggests C. parasitica has been introduced to the UK multiple times over at least two decades through international plant trade.     

解淀粉芽胞杆菌YU-1对水稻纹枯病的防治作用

本文旨在筛选对水稻纹枯病有生防作用的菌株,并初步探索其生防作用机理。收集水稻、甘蓝、黄瓜等不同植物根际土壤,采用稀释分离和对峙培养法筛选对水稻纹枯病菌Rhizoctonia solani Kühn有抑菌作用的菌株;通过离体接种防效、盆栽防效、抑菌谱、对水稻纹枯病菌菌核萌发及形成的抑制作用等方面评价其生防潜力,并对生防菌株进行鉴定。结果表明,从采集的37份根际土壤中共分离获得细菌297株,其中4株对纹枯病菌具有较好的拮抗作用,菌株YU-1对水稻纹枯病菌菌丝生长的抑制率达89.8%;对西瓜枯萎病菌、草莓灰霉病菌、番茄早疫病菌、油菜菌核病菌、小麦赤霉病菌菌丝生长的抑制率均在85%以上;对水稻纹枯病的离体和盆栽防效分别为73.1%和66.3%;对水稻纹枯病菌菌核萌发的抑菌率在92%以上;经生理生化和分子鉴定为解淀粉芽胞杆菌Bacillus amyloliquefaciens。由此可看出,菌株YU-1对水稻纹枯病的防治具有较强的应用价值,具有进一步开发成生物农药的潜力。     

陇南核桃病虫害无公害综合防治策略

针对陇南核桃栽培中病虫害发生问题进行分析论述,提出无公害综合防治策略,对陇南生态环境保护及绿色无公害核桃生产有一定指导意义。     

小麦赤霉病自动监测预警系统应用效果评价

2018年在江苏、陕西、河南、湖北、安徽共18个县(市)安装了小麦赤霉病预报器,在周边设置未防治麦田进行赤霉病调查,并与预警软件平台预测结果相比较,评价小麦赤霉自动监测预警系统的准确性。评价结果表明,2018年该系统预测的准确性达71.8%。结合陕西省植保总站、西安市植保站、渭南华州区植保站、商洛洛南县植保站、安徽凤台县植保站2016年-2018年对该系统的评价结果,证实该系统预测准确性较高,系统工作稳定,自动化程度高,可为小麦赤霉病的科学防控提供重要的参考依据,具有一定的应用前景。     

亚硝基谷胱甘肽还原酶 GSNOR1正调控植物对疫霉菌的抗性

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。     

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

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。     

马铃薯AP2/ERF转录因子的克隆及植物表达载体构建

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。