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。     

A rapid extensigraph protocol for measuring dough viscoelasticity and mixing requirement

The extensigraph is particularly useful in characterizing dough viscoelastic properties; however, testing throughput for standard method is low due to the prerequisite for farinograph water absorption, long dough resting and milling to prepare large amounts of flour. Therefore, a rapid extensigraph method was developed that reduced sample size (165 g wheat) for milling and more than tripled throughput. Wheat is milled in Quadrumat Junior mill with a modified sieving system. The resulting flour (100 g) was mixed with a pin mixer at constant water absorption to allow the evaluation of wheat genotypes at the absorption level they are expected to perform. Dough was subsequently stretched by an extensigraph after 15 min of floor time and 30 min resting. Strong correlations for extensigram R_max (r > 0.93), extensibility (r > 0.64) and area (r > 0.88) were found for the proposed method compared to the standard method. Mixing parameters (time and energy) obtained during dough preparation provided further information about dough strength and mixing requirement. By significantly reducing sample size requirement and increasing testing throughput, this rapid extensigraph method can be widely adopted in milling and baking industry and meets the need for a fast evaluation of dough strength in breeding trials.     

Kernel vitreousness and protein content: Relationship,interaction and synergistic effects on durum wheat quality

Four sets of durum samples were used in this study to further understand the interrelationships among hard vitreous kernels (HVK), protein content, and pigment concentration, with a focus on the interaction and synergistic effects of protein content and vitreousness on durum quality. HVK level increases with higher protein content in the range of 9.5–12.5%, but this relationship is less evident in durum samples with high protein content (12.5–14.5%). Both protein content and kernel vitreousness can significantly affect durum milling quality. White starchy kernels (WSK) in low protein durum have a very detrimental impact on milling and pasta processing quality, but high protein content can mitigate the adverse impact of WSK on durum quality. Although protein content plays a dominant role, higher HVK might contribute positively to pasta firmness. There was no significant difference in yellow pigment content between HVK and WSK. However, pigment loss from semolina to dough was higher for WSK than HVK. Despite the difference in protein content, HVK and WSK have little difference in gluten strength. The monomeric protein was preferentially accumulated in HVK. The glutenin proteins of HVK and WSK were similar in the ratios of 1Bx/1By and HMW/LMW-GS.     

A new lean no time test baking method with improved discriminating power

In response to customer concerns related to gluten strength in commercial baking, the Canadian Grain Commission assessed whether the Canadian Short Process (CSP) test bake method was generating useful data related to intrinsic strength of wheat varieties. Assessment of CSP loaf volume data for Canadian variety trials spanning 2003 to 2013 showed very little correlation with dough strength parameters as measured by farinograph and extensigraph. A lean no time (LNT) test baking method was developed that can better discriminate genotypes and provide objective indicators of the effect of intrinsic dough strength on baking quality. From early method development, through method validation and verification using diverse sets of samples targeting different Canadian wheat classes and grown in three different crop years, results showed the LNT method to be more discriminating and easily adopted by other laboratories. In 2015, the LNT method was adopted as the method of choice in future Canadian variety registration trials. The LNT method is fast, simple and well-suited to high throughput test baking conditions encountered in the evaluation of large numbers of breeder lines. A new objective parameter, loaf top ratio, was also introduced and found to correlate well with dough strength and dough handling properties.     

山葡萄VaCIPK18原核表达与多克隆抗体制备

类钙调磷酸酶B亚基蛋白(CBLs)互作蛋白(CIPKs) 作为类丝氨酸/苏氨酸蛋白激酶,在植物响应非生物胁迫信号转导中起着重要作用。基于前期山葡萄响应低温胁迫转录组测序结果,发现低温胁迫引发的早期伤害及感知阶段的激酶基因中涉及CBL-CIPK 信号通路的VaCIPK18表达显著上调。为进一步研究山葡萄(Vitis amurensis)VaCIPK18激酶参与低温胁迫的功能,采用同源克隆获得了VaCIPK18基因,其开放阅读框为1 320 bp,编码439个氨基酸。基于对VaCIPK18蛋白生物信息学分析,获取胞外结构域中抗原表位丰富的肽段,并将其C端调控结构域(230~439 aa)构建到原核表达载体pET28a-SUMO。将重组表达载体转化至大肠杆菌(E. coli Rosetta)中,经0.8 mmol·L^-1 IPTG、37℃诱导4 h表达出大小为42 kDa的包涵体蛋白。将重组蛋白作为抗原免疫日本大耳白兔,获得anti-VaCIPK18多克隆抗体,经检测具有高效价及特异性。Western Blot结果表明,该抗体可以与葡萄内源CIPK18特异性结合,且在50 kDa位置出现与预期一致的条带。同时,CIPK18在低温胁迫后葡萄叶片中蛋白表达水平与室温下相一致,但两种状态下均存在可能的磷酸化与泛素化修饰现象。本研究结果为进一步探究VaCIPK18的蛋白定位、表达及其功能奠定了基础。     

Generation of mouse monoclonal antibodies specific to tilapia immunoglobulin using fish immunoglobulin/BSA complex for monitoring of the immune response in Nile tilapia Oreochromis niloticus

The simple immunoprecipitation method was used to isolate tilapia immunoglobulin (Ig) for immunization in order to produce monoclonal antibodies (MAbs) specific to tilapia Ig. First, the tilapia antiserum against bovine serum albumin (BSA) was prepared by peritoneal injection of BSA into tilapia, and the tilapia anti‐BSA antiserum was used to precipitate BSA to form the Ig/BSA immune complex. The Ig/BSA immune complex was then injected into Swiss mice for hybridoma production. After fusion, three hybridoma clones producing MAbs specific to the tilapia antibody were selected by dot blot and Western blot. All MAbs (101A, 59G, and 11A) were bound specifically to the heavy chain of immunoglobulin M (IgM). The MAbs 101A and 59G demonstrated twofold higher affinity than MAb 11A and the commercialized antibody. However, MAbs 11A could also bind to the heavy chain of IgM in Asian seabass, Lates calcarifer, as well. These MAbs can be used to monitor the immune responses of individual fish by indirect ELISA upon exposure to various antigens.     

吉林省西部地区滴灌条件下宜机械粒收玉米品种筛选

确定宜机械粒收的玉米品种是构建吉林省西部旱作补灌区玉米丰产增效技术模式的重要内容之一。2018年和2019年在吉林省西部地区开展滴灌条件下宜机械粒收玉米品种的筛选试验,测定38个玉米品种的收获子粒含水率和产量,并按玉米子粒含水率和产量水平采用双向平均法作图进行品种分类。结果表明,本研究初步筛选出5个滴灌条件下宜机械粒收玉米品种,分别是迪卡159、福莱77,稷秾108、吉农大889和优迪919。根据品种综合性状分析,吉农大889较优,其次为迪卡159,可以作为适宜吉林省西部滴灌条件下丰产高效栽培的推荐品种。     

西方语境的中国古典园林审美认知历史溯源

中国古典园林审美不仅仅在中国文化体系中进行讨论，随着全球化进程，西方语境学者也逐渐涉及相关的内容。尝试寻找一种跨文化的“他者”视角，梳理近3个世纪以来西方语境的中国古典园林审美认知经历的3个主要阶段。以18世纪为开端，经过19—20世纪借助西译艺术著作和海外园林建设实现对审美理解从印象到真实的转变，到进入21世纪后诞生的跨学科的诸多新观，这个过程中展现出以中国传统园林审美为代表的打破自身线性历史的闭关自守，回归全球语境的未来中国古典园林研究趋势。     

西汉农业诏书探析简

西汉农业诏书及其所反映的农业政策可分为六个时期。农政主要包括:颁布劝农诏,鼓励农业生产;减免租赋,与民休息;振贷贫民,促进农业生产的恢复;皇帝倡导节俭,躬耕藉田,以劝农天下;建立完善的农业管理体系等内容。这些政策促进了国家农本思想形成、农业技术推广,以及农业经济繁荣,并成为封建社会农政的基本模式,使以农立国的传统正式形成。