Pathogenicity of Rice Blast Fungus Magnaporthe oryzae on Brachypodium distachyon

Inoculation methods for rice blast fungus Magnaporthe oryzae to Brachypodium distachyon were developed to investigate the infection process and symptom development in comparison with those on rice (Oryza sativa) and barley (Hordeum vulgare).M.oryzae could infect leaves,sheathes,stems and panicles of B.distachyon and cause blast disease.Spraying conidial suspension on either intact seedlings or leaf segments induced typical symptoms on B.distachyon.During the intact seedling inoculation,the symptom developed on B.distachyon leaves closely resembled that on rice;but the lesions on B.distachyon had better uniformity in shapes and sizes than those on rice or barley.In the leaf segments inoculation,only initial and low-developed lesions could be found on rice,while normal symptoms on B.distachyon and barley.Inoculated with low-virulent mutants of M.oryzae,B.distachyon produced low-level symptoms.The symptom level of each mutant on B.distachyon corresponded well to that on rice.In addition,typical infection processes presented on B.distachyon leaves:forming melanized appressoria,penetrating into host epidermis and then forming hyphae in epidermal cells.According to these results,B.distachyon can be used as a candidate for studying fungus-plant interactions and as a probable source of disease resistance.     

新的禾本科模式植物——二穗短柄草

水稻（Oryza sativa）是禾本科中目前唯一已完成基因组测序的模式植物,但由于它很早从冷季型禾本科中进化分离出来,故其作为禾本科其他亚科的模式植物有一定局限性,因此,需要一种新的、更适合的模式植物。二穗短柄草（Brachypodium distachyon）是一种温带禾草,归属禾本科早熟禾亚科短柄草族短柄草属,具有基因组小、株型小、自花授粉、一年生、生命周期短、有二倍体和一系列多倍体、易培植、易转化等诸多优点,尤其是与很多重要的谷类作物如小麦、大麦、燕麦、玉米、水稻、高梁等以及许多牧草与草坪草有较近的亲缘关系,因而是温带禾本科植物功能基因纽学以及生物能源作物（如柳枝稷）研究的理想模式植物,作者例举二穗短柄草之所以成为新模式植物系统的依据,并介绍其在遗传进化、基因组特点、功能基因组及形态特征等方面的研究进展。     

二穗短柄草CCCH型锌指蛋白基因家族成员的鉴定和分析

CCCH型锌指蛋白基因以基因家族的形式存在,在信号转导调控、形态发生和逆境应答等诸多生物学过程中起着重要作用。为给该基因的克隆及功能研究提供参考信息,本研究针对二穗短柄草的基因组序列,运用多种生物信息学方法在全基因组范围对CCCH型锌指蛋白基因进行了鉴定分析。结果表明,二穗短柄草共有61个含CCCH基序的蛋白质,它们的基因 DNA序列全长825~13 288 bp,内含子数量0~13个,编码区全长498~4 944 bp,编码165~1 007个氨基酸;共检测到145个CCCH基序,以C X₈ C X₅ C X₃ H和C X₇ C X₅ C X₃ H最为常见;基因不均等地分布在5条染色体上,其中2号染色体分布最多,有21个,5号染色体分布最少,仅4个;系统发育分析表明,61个成员可分为8组。     

二穗短柄草BdGPX4基因的克隆及酶学性质

为解析模式植物二穗短柄草中谷胱甘肽过氧化物酶(Glutathione peroxidase,GPX)在抗氧化过程中的作用机制,对二穗短柄草BdGPX4基因进行克隆,获得681bp的cDNA序列,该基因编码226个氨基酸残基的蛋白质,预测相对分子量是24.49ku。通过亚细胞定位预测,BdGPX4蛋白位于叶绿体和线粒体中。生物信息学分析发现BdGPX4基因启动子具有植物激素、光信号、干旱以及冷胁迫响应元件。结合二穗短柄草基因芯片数据和RT-PCR表达检测结果,发现BdGPX4基因在根、茎、叶、叶鞘、苗芽、穗状花序和种子等部位均表达,在光合组织和生长旺盛部位的表达显著高于非光合和衰老部位,并在根部受到SA诱导表达上调。成功构建BdGPX4基因的原核表达载体,在大肠杆菌中表达并纯化BdGPX4蛋白,检测到对H_2O_2和COOH具有较高活性,但对底物tBOOH活性较低,与直系同源蛋白OsGPX4相比功能发生分化。基因表达模式及酶学活性的特点预示着二穗短柄草BdGPX4基因在光合器官和胁迫条件下具有重要的抗氧化作用。     

The Brachypodium distachyon UGT Bradi5gUGT03300 confers type II fusarium head blight resistance in wheat

Fusarium head blight (FHB), caused by fungi belonging to the Fusarium genus, is a widespread disease of wheat (Triticum aestivum) and other small-grain cereal crops. The main causal agent of FHB, Fusarium graminearum, produces mycotoxins mainly belonging to type B trichothecenes, such as deoxynivalenol (DON), that can negatively affect humans, animals and plants. DON detoxification, mainly through glucosylation into DON-3-O-glucose, has been correlated with resistance to FHB. A UDP-glucosyltransferase from the model cereal species Brachypodium distachyon has been shown to confer resistance both to initial infection and to spike colonization (type I and type II resistances, respectively). Here, the functional characterization of transgenic wheat lines expressing the Bradi5g03300 UGT gene are described. The results show that, following inoculation with the fungal pathogen, these lines exhibit a high level of type II resistance and a strong reduction of mycotoxin content. In contrast, type I resistance was only weakly observed, although previously seen in B. distachyon, suggesting the involvement of additional host-specific characteristics in type I resistance. This study contributes to the understanding of the functional relationship between DON glucosylation and FHB resistance in wheat.     

二穗短柄草SPL家族基因的鉴定、系统发育和表达分析

植物特异性SPL家族基因编码SBP（squamosa promoter-binding protein-like）保守结构，参与调节植物的生长和发育。为了解二穗短柄草中SPL家族基因的分布、结构及功能，对二穗短柄草全基因组中SPL家族成员进行了系统分析。结果表明，在二穗短柄草全基因组中鉴定出17个BdSPL基因，在5条染色体上呈不均匀分布,bd03号染色体上分布最多；根据系统发育树，17个BdSPL被分为6个组，同组成员有保守的结构和基序；其中6对 BdSPL基因发生基因复制事件;在BdSPL基因启动子区域发现了各种顺式元件，如ABRE、CGTAC-motif和LTR。转录组数据和qRT-PCR分析显示，BdSPL基因在二穗短柄草花中表达量较高，而在根系中表达量较低，在不同植物激素处理下表达量有差异。     

Can wild species become problem weeds because of herbicide resistance? Brachypodium distachyon: a case study

A purported drawback to the use of transgenic herbicide-resistant crops is the fear that the crop or interbreeding wild relatives will become weedy. It has been posited that a change even in a single trait can confer weediness. This hypothesis was tested with Brachypodium distachyon. This innocuous species came into contact with herbicides through the use of crushed rock from its habitat for road foundations. It evolved s-triazine resistance and developed as a monoculture. When true weeds later evolved simazine resistance, B. distachyon was partially competed from the ecosystem and then disappeared upon the use of non-triazine herbicides. Thus, this wild species remained a weed only until true weeds evolved resistance or until other herbicides were used. One gene mutation did not convert it into a weed, which implies that this will be equally improbable in other cases, when the gene codes for an otherwise neutral trait such as herbicide resistance.     

二穗短柄草GRFs基因家族的鉴定及表达模式分析

GRFs基因家族在植物生长发育及逆境响应中起着重要的调控作用。为了探明二穗短柄草GRFs基因家族的基本特征及其进化关系,本研究利用生物信息学方法在全基因组水平上对二穗短柄草GRFs基因家族成员进行鉴定与分析。结果表明,二穗短柄草GRFs基因家族包括12个家族成员,蛋白质序列长度在238~577个氨基酸之间;序列比对发现二穗短柄草GRFs家族基因包括2个保守的QLQ和WRC结构域及广泛的保守基序;染色体定位发现,12个家族成员在二穗短柄草的5条染色体上呈不均匀分布,bd03号染色体分布最多;系统发育关系比较分析表明,这些GRFs基因家族成员存在4对直系同源基因。RT-qPCR分析表明,全部GRFs基因家族成员在二穗短柄草幼嫩组织中表达量较高,而在根系和衰老组织中表达量较低,外源激素特别是GA₃诱导了二穗短柄草大部分GRFs基因家族成员的上调表达,表明GRFs基因家族广泛地参与了二穗短柄草分生组织功能和器官形成的生命进程。本研究结果为二穗短柄草GRFs家族蛋白功能的研究奠定了理论基础。     

二穗短柄草2C型蛋白磷酸基因BdPP2C1的克隆及表达分析

从二穗短柄草中扩增得到一个2C型蛋白磷酸酶基因,命名为BdPP2C1。BdPP2C1与拟南芥中PP2C家族进行系统进化分析,结果表明,BdPP2C1与拟南芥A类PP2C家族成员的亲缘关系较近。利用实时荧光定量PCR表达分析系统,分析在非生物逆境胁迫及相关信号分子处理下BdPP2C1基因的表达情况。结果表明:该基因的表达受脱落酸(ABA)和双氧水抑制,并且受渗透和低温胁迫诱导。因此,BdPP2C1是非生物逆境胁迫中的一个应答因子,并且可能受相关信号分子调控。