Calmodulin interacts with MLO protein to regulate defence against mildew in barley

In plants, defence against specific isolates of a pathogen can be triggered by the presence of a corresponding race-specific resistance gene, whereas resistance of a more broad-spectrum nature can result from recessive, presumably loss-of-regulatory-function, mutations. An example of the latter are mlo mutations in barley, which have been successful in agriculture for the control of powdery mildew fungus (Blumeria graminis f. sp. hordei; Bgh). MLO protein resides in the plasma membrane, has seven transmembrane domains, and is the prototype of a sequence-diversified family unique to plants, reminiscent of the seven-transmembrane receptors in fungi and animals. In animals, these are known as G-protein-coupled receptors and exist in three main families, lacking sequence similarity, that are thought to be an example of molecular convergence. MLO seems to function independently of heterotrimeric G proteins. We have identified a domain in MLO that mediates a Ca2+-dependent interaction with calmodulin in vitro. Loss of calmodulin binding halves the ability of MLO to negatively regulate defence against powdery mildew in vivo. We propose a sensor role for MLO in the modulation of defence reactions.     

Processing of wild cereal grains in the Upper Palaeolithic revealed by starch grain analysis

Barley (Hordeum vulgare L.) and wheat (Triticum monococcum L. and Triticum turgidum L.) were among the principal 'founder crops' of southwest Asian agriculture. Two issues that were central to the cultural transition from foraging to food production are poorly understood. They are the dates at which human groups began to routinely exploit wild varieties of wheat and barley, and when foragers first utilized technologies to pound and grind the hard, fibrous seeds of these and other plants to turn them into easily digestible foodstuffs. Here we report the earliest direct evidence for human processing of grass seeds, including barley and possibly wheat, in the form of starch grains recovered from a ground stone artefact from the Upper Palaeolithic site of Ohalo II in Israel. Associated evidence for an oven-like hearth was also found at this site, suggesting that dough made from grain flour was baked. Our data indicate that routine processing of a selected group of wild cereals, combined with effective methods of cooking ground seeds, were practiced at least 12,000 years before their domestication in southwest Asia.     

大麦醇溶蛋白基因片段克隆和RNAi植物表达载体构建

 为了降低啤酒大麦蛋白的含量,提高啤酒大麦品质,利用甘肃推广面积最大的品种甘啤4号为材料,克隆大麦醇溶蛋白基因保守区序列,构建RNAi表达载体。根据已知大麦醇溶蛋白基因保守序列设计一对特异性引物B2PF5′-CAACCATTTCCACAGCAACCACCAT-3′,B2PR5′-GAAAGATAGAGTAGACGATTGCACG-3′,采用PCR技术从甘啤4号大麦基因组中获得了1个349bp片段（GP4）。依据载体pHANNIBAL和pART27的结构,分别将GP4按所对应酶切位点正反向插入,利用热激法转化。结果分析显示,克隆出的片段与GenBank （NCBI）中醇溶蛋白基因核苷酸序列同源性高达92%,所构建成的RNAi表达载体pART27-pHAN-GP4中含目标片段。     

SNARE-protein-mediated disease resistance at the plant cell wall

Failure of pathogenic fungi to breach the plant cell wall constitutes a major component of immunity of non-host plant species--species outside the pathogen host range--and accounts for a proportion of aborted infection attempts on 'susceptible' host plants (basal resistance). Neither form of penetration resistance is understood at the molecular level. We developed a screen for penetration (pen) mutants of Arabidopsis, which are disabled in non-host penetration resistance against barley powdery mildew, Blumeria graminis f. sp. hordei, and we isolated the PEN1 gene. We also isolated barley ROR2 (ref. 2), which is required for basal penetration resistance against B. g. hordei. The genes encode functionally homologous syntaxins, demonstrating a mechanistic link between non-host resistance and basal penetration resistance in monocotyledons and dicotyledons. We show that resistance in barley requires a SNAP-25 (synaptosome-associated protein, molecular mass 25 kDa) homologue capable of forming a binary SNAP receptor (SNARE) complex with ROR2. Genetic control of vesicle behaviour at penetration sites, and plasma membrane location of PEN1/ROR2, is consistent with a proposed involvement of SNARE-complex-mediated exocytosis and/or homotypic vesicle fusion events in resistance. Functions associated with SNARE-dependent penetration resistance are dispensable for immunity mediated by race-specific resistance (R) genes, highlighting fundamental differences between these two resistance forms.     

4个小麦品种的抗白粉病遗传分析

抗病品种郑315、CP87-26-12和91138-16-2-13-12,与感病品种豫麦49号杂交的F1代表现抗病,F2代抗、感单株的分离比例为31,表明这3个品种均携带1对显性抗病基因.N97189-2和豫麦18号的杂交F1代对白粉病表现高感,F2代抗、感单株的分离比例为13,由此推断N97189-2对白粉病的抗性由1对隐性基因控制.     

一个烟草Mlo基因的电子克隆及其序列特性分析

电子克隆并探讨了一个烟草Mlo基因的结构和功能．以GenBank公布的水稻Mlo基因序列为信息探针对烟草的EST数据库进行同源搜索,并将获得的同源性高的EST序列进行序列拼接,最后得到了一个烟草Mlo基因的cDNA序列．采用生物信息学方法分析该基因编码蛋白的一级、二级结构,并对其功能进行预测．结果表明,此烟草Mlo基因...     

VIGS技术初步鉴定RgMLO6和RlMLO7基因对白粉病的负调控功能

为了探讨蔷薇科植物MLO基因在抗白粉病中的作用,研究应用病毒诱导的基因沉默技术(virus induced gene silencing,VIGS)抑制了大花香水月季RgMLO6基因和长尖叶蔷薇RlMLO7基因的表达,随后接种白粉菌对这2个基因进行抗性鉴定. 研究发现在VIGS载体转化植株叶片20 d后,RgMLO6和RlMLO7基因的相对表达量显著下降了80%～90%,沉默效果明显. 分别对2个基因沉默后的嫩叶进行白粉病抗性鉴定,大花香水月季和长尖叶蔷薇的抗性水平较对照组均提高. 显微镜观察白粉菌接种2个基因沉默后植株叶片中菌丝体的生长情况,整体表现出沉默植株叶表皮细胞上的白粉菌生长较对照组生长缓慢. 结果表明RgMLO6与RlMLO7基因对蔷薇科植物的白粉病有负向调控作用.     

用RT-PCR技术研究水稻中抗病基因同源序列DFR-1、OsMlo-1所在基因的功能

DFR-1、OsMlo-1分别是最近从水稻中克隆的玉米Hml和大麦中Mlo抗病基因同源序列，这两个序列与两个已报道的水稻抗稻瘟病数量性状位点（QTLs）有较好的对应关系，表明它们所在的基因可能参与抗病反应，为了进一步研究水稻DFR-1、OsMlo-1所在基因的功能，在DFR-1、OsMlo-1假定的外显子上设计引物，通过RT-PCR技术，研究在接种白叶枯病（Xanthomanas oryza pv oryzae）菌株PX099以及接种稻瘟病（Magnoparthe grisea）菌株V86013前后，水稻品种IRBB13和水稻品种明恢63中DFR-1、OsMlo-1所在基因的表达．结果表明：在接种白叶枯病菌株PX099的水稻品种IRBBl3中与DFR-1对应的基因是诱导增强的；在接种稻瘟病菌株V86013的水稻品种明恢63中与DFR-1、OsMlo-1对应的基因是诱导增强的，进一步表明DFR-1、OsMlo-1所在的基因可能参与水稻抗病反应。     

黄化型大麦的系列研究 Ⅰ南黄大麦的发现

南黄大麦系辐射突变选育而成。植株前期叶色浅黄,后期淡绿,是一种稀有的黄化型大麦。南黄大麦的发现,不仅丰富了大麦的基因库,而且在遗传、核质关系,超微结构、光合作用等方面具有理论和实践的意义。     

Mobilization of a transposon in the rice genome

Rice (Oryza sativa L.) is an important crop worldwide and, with the availability of the draft sequence, a useful model for analysing the genome structure of grasses. To practice efficient rice breeding through genetic engineering techniques, it is important to identify the economically important genes in this crop. The use of mobile transposons as gene tags in intact plants is a powerful tool for functional analysis because transposon insertions often inactivate genes. Here we identify an active rice transposon named miniature Ping (mPing) through analysis of the mutability of a slender mutation of the glume-the seed structure that encloses and determines the shape of the grain. The mPing transposon is inserted in the slender glume (slg) mutant allele but not in the wild-type allele. Search of the O. sativa variety Nipponbare genome identified 34 sequences with high nucleotide similarity to mPing, indicating that mPing constitutes a family of transposon elements. Excision of mPing from slg plants results in reversion to a wild-type phenotype. The mobility of the transposon mPing in intact rice plants represents a useful alternative tool for the functional analysis of rice genes.     

用基因芯片技术分析干旱胁迫下生殖生长期大麦的基因表达

耐旱性是干旱地区稳定和增加大麦产量的一个关键因素。鉴定出与耐旱性相关的功能基因,一方面可了解大麦的耐旱机理,同时还可以促进利用生物技术来改良大麦的耐旱性。在研究中,2个在耐旱性上具有明显差异的大麦品种Tadmor(耐旱)和WI2291(干旱敏感)被选作材料,采用22000个ESTs(基因表达序列标签)的Affymetrix大麦基因芯片Barley1来分析生殖生长期干旱胁迫下2个大麦材料的差异表达基因。研究结果表明,干旱胁迫下2个大麦材料中有77个共调节基因,其中部分基因已被报道过可能与抗旱性相关。这些基因中已有功能注释的基因按其生物学功能被分为14组,猜测它们是干旱胁迫的响应基因,在抗旱性上可能不起重要作用,或者是必需的但单独不足以提高大麦的抗旱性。进一步比较2个材料差异表达的基因,发现二材料之间有372个受干旱调节基因的差异。这些基因中有功能注释基因的生物学功能中可分为15组,其中一些已被认为与抗旱性相关;而对那些未知功能的基因,推测可能亦在大麦的抗旱性上扮演一定的角色。研究所得结果可为阐述生殖生长期大麦的耐旱性机理提供新的认识。     

小麦中源于中间偃麦草抗白粉病基因PmCH5026的SSR定位

CH5026是衍生于八倍体小偃麦TAI7045的新品系,用高感品种(系)CH5065、晋太170分别与CH5026配置组合,于温室接种并调查F2、F3、BC1F2群体的抗感分离之比进行遗传分析,结果表明CH5026成株期对白粉菌E09小种的抗性受1对显性基因控制,暂命名为PmCH5026.使用集群分离分析法(BSA),用378对SSR引物对CH5026×CH5065 F2代群体进行分析,筛选到标记Xcfd233、Xbarc11和Xgwm539与抗性基因连锁,位置顺序为:Xcfd233-7.2cM-PmCH5026-4.9cM-Xbarc11-5.5cM-Xgwm539.根据小麦微卫星遗传连锁图及利用中国春第2同源群缺四体、双端体对SSR标记的定位结果,将PmCH5026定位在染色体2DL上.     

转多个抗真菌蛋白基因水稻植株的获得及其抗稻瘟病菌的初步研究

用基因枪法将水稻碱性几丁质酶（ＲＣ２４）基因、苜蓿葡聚糖酶（βＧｌｕ）基因、大麦核糖体失活蛋白（ＢＲＩＰ）基因和潮霉素（ｈｐｔ）基因同时导入籼稻品种（七丝软占）中,获得了７个潮霉素抗性再生系,Ｓｏｕｔｈｅｒｎｂｌｏｔ证明２～３个抗真菌蛋白基因已整合到水稻基因组中．初步抗性鉴定表明Ｒ０代转基因水稻植株对稻瘟病菌的抗性有所提高     

A transgenic wheat with a stilbene synthase gene resistant to powdery mildew obtained by biolistic method

Stilbene, a kind of phytoalexin, plays an important role in resistance to fungal and bacterial infection in plants. It strongly inhibits the growth of fungi and sprout of spore. Stilbene synthase gene (Vst1) obtained from grapevine has been transferred into common spring wheat Jinghong 5 by using the biolistic transformation method. Five transgenic plants (T₀) were obtained from the bombarded 2014 immature embryos. One immune plantlet and 3 plantlets with mid-resistance to powdery mildew were identified from the transgenic plants of T₃ generation which came from 2 T₀ transgenic plants.     

短柄五加(Acanthopanax brachypus)rbcL基因的结构分析

克隆了含完整短柄五加rbcL基因的3.2kb EcoRI片段,测定了该基因的核苷酸序列.所测核苷酸序列总长度为1924bp,其中编码区1428bp,编码475个氨基酸的蛋白质.测定的基因5’上游区共278bp,包含原核性质-35区(TTGCGC),-10区(TACAAT)及类似真核的TATA box元件(TATATA).5’前导区长194bp,其中SD序列为GGAGG,紧邻起始密码子上游.测定的3’下游区共218bp,含2个相邻的转录后可形成茎环结构的反向重复序列.短柄五加rbcL基因编码区推导的氨基酸序列与烟草、菠菜、豌豆、苜蓿、玉米、水稻、松树、地钱、衣藻和Anacystis的同源性分别为93.5％、94.11％、94.53％、94.74％、89.68％、92.21％、92.21％、92.63％、87.58％和80.84％.本文还对不同植物rbcL基因的启动区及部分5’和3’非编码区进行了比较分析.     

Familial combined hyperlipidaemia linked to the apolipoprotein AI-CII-AIV gene cluster on chromosome 11q23-q24.

Familial combined hyperlipidaemia (FCHL) is a common inherited disorder of lipid metabolism with a prevalence of 0.5-2.0% (refs 1, 2). It is estimated to cause 10% of premature coronary heart disease. The underlying metabolic and genetic defects in FCHL have not been identified, but a population study has suggested an association between FCHL and an XmnI restriction fragment length polymorphism (RFLP) within the apolipoprotein AI-CIII-AIV gene cluster. Here we confirm this association and show that it results from linkage disequilibrium between FCHL and the 6.6-kilobase (kb) allele of the XmnI RFLP. Subsequent analysis in seven FCHL families, ascertained through a proband carrying the 6.6 kb XmnI allele, demonstrated linkage to the AI-CIII-AIV cluster on 11q23-q24, zeta = 6.86 with no recombinants. This assignment will facilitate the identification of the mutation that causes hyperlipidaemia in these families.     

Breeding and molecular cytogenetic identification of wheat-Thinopyrum intermedium addition lines resistant to powdery mildew

Wheat-related species Th. intermedium was used to cross with common wheat Yannong 15. In the self progenies of the hybrid, two addition lines, Ⅱ-1-7-1 and Ⅱ-3-3-2, stable in cytology, were developed by cytology and powdery mildew resistance identification. Their chromosome number were 2n = 44 and formed 22 bivalents at PMC MI. In F1 of the two addition lines crossing with Yannong 15, there appeared about one univalent at PMC MI, respectively. Resistance identification in greenhouse and field using the No. 15 and mixed strains of E. gramnis f. sp. tritici showed that they were immune to powdery mildew. Chromosome number and resistance identification using the F2 single plants of the addition line crossing with Yannong 15 indicated that the resistant gene was located on the alien chromosomes. In situ hybridization using St and E genomic DNA as probe showed that the added chromosome in the two addition lines probably came from the E genome of Th. intermedium, which indicated that a pair of E genome chromosomes carried a new resistant gene to powdery mildew.     

Obtained transgenic wheat expressing pac1 mediated by Agrobacterium is resistant against Barley yellow dwarf virus-GPV

In fission yeast （Schizosaccharomyces pombe）, pacl gene was cloned with 99.3% nucleotide sequence similarity with published pacl in GenBank. In pET-5α expression system, the expression product of cloned pacl in E. coil showed activity to degrade the double-strand RNA. Harboring the binary vector pBI121, which contains pacl gene, Agrobacterium tumefaciens strain LBA4404 was used to transform the wheat immature embryos precultured 7-10 d. After preregeneration, regeneration and selection culture stage, totally 41 G418 resistant plants were obtained, in which 25 lines were proved to integrate with transgene and express transgene normally by PCR, Dot blot, RT-PCR and ELISA detection. Antivirus test carried out on 25 positive lines with high dose of Barley yellow dwarf virus-GPV revealed that 12 lines had resistance to BVDV-GPV in low level, another 12 lines had resistance to BVDV- GPV in middle level, and 1 line showed resistance to BVDV-GPV in high level. However, both low and middle level of resistance plants showed no symptoms when infected by viruses at low dose, which suggested the dose-dependent effect of the resistance mediated by pacl to BYDV-GPV.     

大麦及其制品质量安全风险及控制

大麦是我国进口量和进口额最大的谷物品种。近半个世纪以来,世界大麦生产和需求呈持续增长的趋势,全球贸易日趋活跃。概述了大麦的营养及我国大麦产业发展概况,指出大麦及其制品可能存在质量安全问题的来源,提出保障大麦及其制品质量安全的调控建议,以期为我国大麦产业健康快速发展提供一些思考和借鉴。     

RNA toxicity is a component of ataxin-3 degeneration in Drosophila

Polyglutamine (polyQ) diseases are a class of dominantly inherited neurodegenerative disorders caused by the expansion of a CAG repeat encoding glutamine within the coding region of the respective genes. The molecular and cellular pathways underlying polyQ-induced neurodegeneration are the focus of much research, and it is widely considered that toxic activities of the protein, resulting from the abnormally long polyQ tract, cause pathogenesis. Here we provide evidence for a pathogenic role of the CAG repeat RNA in polyQ toxicity using Drosophila. In a Drosophila screen for modifiers of polyQ degeneration induced by the spinocerebellar ataxia type 3 (SCA3) protein ataxin-3, we isolated an upregulation allele of muscleblind (mbl), a gene implicated in the RNA toxicity of CUG expansion diseases. Further analysis indicated that there may be a toxic role of the RNA in polyQ-induced degeneration. We tested the role of the RNA by altering the CAG repeat sequence to an interrupted CAACAG repeat within the polyQ-encoding region; this dramatically mitigated toxicity. In addition, expression of an untranslated CAG repeat of pathogenic length conferred neuronal degeneration. These studies reveal a role for the RNA in polyQ toxicity, highlighting common components in RNA-based and polyQ-protein-based trinucleotide repeat expansion diseases.