我国49种线状植物病毒分子鉴定及其基因组研究

本研究建立了马铃薯Y病毒属、马铃薯X病毒属、麝香石竹潜隐病毒属和葱X病毒属成员RT-PCR检测和全基因组扩增技术体系,并成功用于60多种供试植物病毒的检测诊断。利用所建立的技术体系,从全国18个省市42种作物上采集的病样中鉴定了49种植物病毒,其中10种为新种,11种为国内新记录,更正了7种病毒的鉴定和命名。测定了这49种病毒217个分离物基因组序列,并全部在国际基因数据库登录,占全球登录的植物病毒基因组序列总数的3.8%,其中27种植物病毒为基因组全序列,20种为国际首次报道,15种病毒全序列被美国国家生物技术信息中心(NCBI)确定为相关病毒的标准序列。     

Arabidopsis ethylene receptors have different roles in Fumonisin B1-induced cell death

Ethylene is a central signalling agent in mediating plant defence against pathogens. Mutations to the ethylene receptor ETR1 have been shown to alter susceptibility of plants to mycotoxin-induced cell death. Using Fumonisin B₁ (FB₁) to induce cell death, we demonstrate that the receptor mutant ein4-1 has a reduced rate of necrosis, potentially due to an upregulation of ETHYLENE RESPONSE FACTOR1. Mutations in other ethylene receptors differentially affected the expression of genes in the jasmonic and salicylic acid defence pathways. Together these data indicate that ethylene receptors do not have redundant roles in mediating FB₁-induced cell death.     

Possible involvement of gibberellins and ethylene in Orobanche ramosa germination

Summary Six inhibitors acting at different stages of gibberellin biosynthesis, applied during conditioning of Orobanche ramosa seeds, reduced subsequent germination in the presence of GR₂₄ (a strigol analogue). Ethylene seems to be involved in the induction of germination of conditioned seeds by GR₂₄, as inhibitors of its synthesis or action, applied to conditioned seeds, also strongly reduced induction of germination by GR₂₄. Exogenous ethylene did not induce germination of conditioned seeds, but 2-chloroethylphosphonic acid was able to do so. When inhibitors of gibberellin biosynthesis were applied to conditioned seeds in the presence of GR₂₄, they inhibited germination. These same inhibitors also strongly inhibited germination of conditioned Striga hermonthica seeds in response to GR₂₄; this inhibitory effect was reversed by the addition of 2-chloroethylphosphonic acid. The effect of these inhibitors on S. hermonthica , in which ethylene is a necessary mediator of germination induction by GR₂₄, strongly suggests that ethylene synthesis is also required for the induction of O. ramosa seed germination by GR₂₄. These growth regulators, which inhibit the two steps of germination in O. ramosa , could be useful for the development of methods for early season control of this parasite.     

Agrobacterium induces plant cell death in wheat (Triticum aestivum L.)

The symptoms of gall or hairy root do not occur in the interactions between wheat (Triticum aestivum L.) and other monocotyledonous plants, with Agrobacterium tumefaciens or Agrobacterium rhizogenes. However, both bacteria colonized wheat root surfaces at similar levels (2.0 × 10⁷ colony forming U g⁻¹ root) and grew without inhibition in suspension with intact or wounded wheat embryos or root segments present. Suspension-cultured wheat embryo cells grown in 7.4 m M O₂ displayed 23% cell death after 1 h exposure to Agrobacterium cells, while the extent of cell death with 2.1 m M O₂ averaged 8%. Cultured wheat embryo and root cells rapidly produced hydrogen peroxide (H₂O₂) when contacted with A. tumefaciens or A. rhizogenes. Production of H₂O₂ was lower at 2.1 m M O₂ than 7.4 mM O₂. Browning and autofluorescence of epidermal cells of callus derived from wheat embryos and wheat roots was observed after inoculation with Agrobacterium. An increase in ferulic acid was detected in the walls of roots exposed to Agrobacterium. However, neither lignin nor callose was detected by diagnostic staining methods. These findings suggest that Agrobacterium induced a resistance-like response in wheat that may reduce the efficacy of transformation and limit the normal symptom formation.     

The Colletotrichum higginsianum secreted effector protein ChEC91 induces plant cell death

ChEC91, a novel cell death-inducing effector protein from the fungal pathogen Colletotrichum higginsianum, causal agent of crucifer anthracnose disease, is described. Both transient expression of ChEC91 and infiltration of purified recombinant protein induced necrotic lesions in Nicotiana benthamiana leaves. The recombinant protein also induced electrolyte leakage and callose deposition in Arabidopsis thaliana leaf tissue and the expression of defence marker genes. Moreover, fungal mutants constitutively over-expressing ChEC91 in C. higginsianum were impaired in appressorial penetration on Brassica rapa cotyledons. These results suggest that inappropriate expression of ChEC91 might negatively affect the early stage of C. higginsianum infection by inducing plant defence responses. Protein domain deletion analysis showed that the C-terminal region of ChEC91 was necessary, but not sufficient, for activity in N. benthamiana. Homologous effector proteins cloned from C. gloeosporioides, Fusarium graminearum, and Pyricularia oryzae differed in their cell death-inducing activity, which appeared related to sequence variations in the C-terminal region of these proteins. Moreover, this region contained amino acid residues that were well conserved within Colletotrichum species. These results suggest that the amino acid residues in the C-terminal region may be important for inducing cell death in plants.

     

农作物分子遗传图谱的研究进展

概略介绍了分子遗传图谱构建的理论基础及其所需的分子标记和作图群体的种类;重点汇总了已构建的农作物的分子遗传图谱;并就农作物分子遗传图谱构建存在的问题和今后的研究方向进行了讨论。     

Conditional expression of harpinPsscauses yeast cell death that shares features of cell death pathway with harpinPss-mediated plant hypersensitive response (HR)

Conditional expression of harpin_Psscauses yeast cell death that shares features of cell death pathway with harpin_Pss-mediated plant hypersensitive response (HR).Pseudomonas syringae pv.syringae 61 hrp Z gene encodes harpin_Pss, a 34.7 kD extracellular protein that elicits a hypersensitive response (HR) in plants. Conditional expression of either full-length or truncated hrp Z sequences under the GAL1 promoter caused cell death in Saccharomyces cerevisiae Y187. Plating of pYEUT- hrp Z transformants on a medium containing galactose resulted in complete inhibition of colony formation, whereas their growth on a glucose-based medium was unaffected. Western blot analysis confirmed the expression of harpin_Pssin yeast cells transformed with pYEUT- hrp Z and grown in galactose-containing medium. A time-dependent decline in the percentage of trypan blue-excluding cells in cultures of pYEUT- hrp Z transformants was observed when cultured on galactose-containing medium. Similarly, the number of viable cells reduced to about 50% within 6 h. There were similarities in the harpin_Pss-mediated cell death in plants and yeast cell death (YCD). Galactose-induced cell death in pYEUT-hrp Z transformants of S. cerevisiae Y187 was suppressed by a protein kinase inhibitor K252a (10 μ M). The viability of pYEUT- hrp Z transformants was prolonged in the presence of 100 U ml⁻¹catalase suggesting a role for the oxidative burst in YCD that was further supported by the flow cytometric patterns of propidium iodide uptake by yeast cells. Overall, it appears that yeast provides a useful model system to understand the molecular mechanism of harpin_Pss-mediated cell death.     

植物几丁质酶在抗真菌病害基因工程中的应用

植物几丁质酶因能水解真菌细胞壁的主要成分几丁质 ,在抗植物真菌病害反应中发挥了重要的作用。本文对植物几丁质酶的特性、分类、基因的诱导表达及其基因在抗真菌病害基因工程中的应用等进行了阐述 ,指出植物几丁质酶在抗真菌病害基因工程中的巨大应用前景     

The involvement of ethylene and calcium in gray mold of pelargonium,ruscus, and rose plants

Leaves ofPelargonium domesticum andRuscus hypoglossum infected byBotrytis cinerea Pers., produced 12 and 1.5 nl ethylene/h/g, respectively, 34 days after inoculation; wounded or healthy leaves and phyloclades of them produced much lower amounts. When the fungus was grown on dead leaves it produced negligible amounts of the gas even when supplemented with methionine. Exogenous ethylene enhanced gray mold development in both hosts. Silver thiosulfate, aminooxyacetic acid (AOA) and aminoethoxyvinylglycine significantly inhibited disease development in pelargonium, and the latter two compounds inhibited ethylene production. AOA inhibited disease development and ethylene production in cut rose flowers; calcium ions inhibited disease development whereas the chelator EGTA [ethylene glycol bis-(β-aminoethyl ether)N,N,N′,N′-tetraacetic acid] enhanced it. Disease suppression by an excess of Ca²⁺ was correlated with repression of ethylene production by the flowers, whereas deficiency in Ca²⁺ increased disease severity. Publication of the Agricultural Research Organization. No. 2413-E, 1988 series.     

植物与病原物互作中的细胞程序化死亡

细胞程序化死亡(PCD)广泛存在于动植物的生长发育或抗逆过程中,而植物与病原物互作中的PCD已成为当前植物病理学的研究热点之一。本文从植物细胞程序化死亡的概念、检测方法、植物与病原物互作间的细胞程序化死亡及调控机理等方面进行了简要概述。     

Soybean sudden death syndrome: Fungal pathogenesis and plant response

Soybean sudden death syndrome (SDS) is a fungal disease caused by members of clade 2 of the Fusarium solani species complex (FSSC). These fungi are soilborne pathogens that infect soybean plants through the roots and produce toxins that translocate to aerial parts of the plant, inducing foliar chlorosis and necrosis followed by premature defoliation. Here, we first give the current state of knowledge of early pathogen detection and infection establishment for the SDS pathosystem. Subsequently, we discuss the nature and activity of secreted toxins, followed by an overview of changes in plant metabolism and factors that influence fungus–soybean interaction. Finally, we summarize the advances in plant disease resistance, symptom evaluation, and treatment.     

抗昆虫蝎毒素及其在植物遗传转化中的研究进展

本文对抗昆虫蝎毒素的来源、种类、作用机理及其在植物遗传转化方面的研究现状做一简要介绍,并对其在植物遗传转化方面所面临的主要问题进行了探讨,为更好地利用抗昆虫蝎毒素基因提高植物抗虫性提供参考。     

植物凝集素在抗刺吸式害虫转基因工程中的应用

植物凝集素是一类具有高度特异性糖结合活性的蛋白,含有1个或多个可与单糖或寡聚糖特异可逆结合的非催化结构域,在农业生产上应用前景广阔。本文简要概述了几种植物凝集素:雪莲凝集素、天南星科植物凝集素、苋菜类植物凝集素在抗刺吸害虫基因工程上的应用,并简单介绍了转双价抗虫基因方面的研究。     

两种林木植原体病害的分子鉴定

近年来, 林木植原体病害发生日趋严重, 对我国林业经济和生态造成了很大损失。2021年-2022年, 在山西沙棘主产区的沙棘上和北京冬奥园区的油松上分别出现了典型的植原体病害症状;沙棘叶片皱缩, 呈小叶状;油松过度分枝生长且出现球状结构等。本研究通过荧光显微观察, 16S rRNA和tuf基因序列分析, 并结合虚拟限制性片段长度多态性分析证实了这两种病害均由植原体引起。基于16S rRNA的系统进化分析显示:引起沙棘叶片皱缩的植原体与泡桐丛枝植原体(Paulownia witches’-broom phytoplasma, OP107515.1)的相似性最高(99.92%), 引起油松丛枝的植原体与狗尾草丛枝植原体株系(Setaria viridis witches’-broom phytoplasma, FJ263625.1)的相似性最高(99.52%);基于tuf基因的系统发育树显示, 二者同属于16SrⅠ组D亚组(16SrⅠ-D)。本研究首次明确了沙棘叶片皱缩病和油松丛枝病相关植原体的分类地位, 为这两种植原体病害的准确诊断、快速检测及其防治提供了基础资料。