水稻主基因-多基因混合遗传控制白叶枯病抗性的基因效应分析

利用主基因－多基因混合遗传模型分析了５个抗感交组合对水稻白叶枯病菌抗性的基因效应,结果表明５个组合中的３个主基因抗性遗传符合德尔分离比的前提下存在多基因抗性,而且这３个组合彼此间抗病基因的加性效应,主基因和多基因遗传方差及其遗传率存在变异。说明水稻白叶枯病抗性虽以主基因作用为主,但考虑到抗性的持久性,建议在水稻白叶枯病育种中构建主基因－多基因混合遗传体系,以有效抑制白枯病菌群体中小种的波动。     

水稻对白叶枯病Xanthomonas oryzae pv.Oryzae水平抗性的数量性状位点(QTL)定位

利用“Lemont/特青”重组自交系（RI）群体研究了水稻对白叶枯病致病菌株CR6的水平抗性。双亲和F1均为感病,重组自交系（RILs）的病斑长度（LL）为带有明显双向超亲的连续变异,显示出典型的多基因遗传特征。部分重组自交系（约占总数90％）对CR6表现高水平抗性（LL≤3cm）。利用由178个良好分离的RFLP标记构建的饱和连锁图,鉴定出11个数量形状位点（QTLs）和3对互作位点解释了RI群体的大部分病斑变异。抗性QTLs定位于水稻第2、3、4、8、9、10、11、12等8条染色体。在来自特青的Xa－4位点上检测到一个有很大加性效应的QTL。其余10个QTLs的抗性等位基因有7个来自特青,3个来自Lemont。研究结果表明多个数量性状位点和失效主基因（Xa－4）残效的累加效应构成了对白叶枯病水平抗性的遗传基础,是重要的抗性组成部分。可以预期在DNA标记的辅助下,这些数量性状位点与主效抗性基因的组合将使水稻品种具有持久抗病性。     

Culture medium for Xanthomonas campestris pv. oryzae

Y uan , W. 1990. Culture medium for Xanthomonas campestris pv. oryzae. Journal of Applied Bacteriology 69 , 798–805.
Studies on nutrient requirements of four Chinese strains of Xanthomonas campestris pv. oryzae in a modified Watanabe's medium led to the development of a new synthetic medium containing sucrose, sodium glutamate, methionine, KH₂PO₄, NH₄C1 and iron chelated with EDTA. The concentration of each ingredient was optimized based on the number of colonies and time required for their appearance. Various concentrations of some nutrients were compared based upon their effects on growth of the pathogen strains and 34 contaminants from rice materials. Tryp-tone enhanced the growth of X. c. oryzae more than that of many contaminants, including Erwinia herbicola . Peptone stimulated growth of X. c. oryzae without promoting excessive contamination. When compared with other media used for X. c. oryzae , the new culture medium enriched with tryptone and peptone gave the highest recovery and earliest appearance of colonies of Chinese strains of this bacterium.     

Phosphorylation of proteins in Xanthomonas campestris pv. oryzae

Protein phosphorylation was studied in Xanthomonas campestris pv. oryzae in vivo and in vitro. In vitro labelling showed that the protein kinases in this bacterium used both ATP and GTP as nucleotide substrates at nearly the same efficiency. At least 6 proteins were phosphorylated in vitro, including abundant species of p81, p44, and p32 with M _r of 81000, 44000, and 32000, respectively. Three types of phosphate-protein linkage were found in this bacterium: O-phosphate, N-phosphate and probably acyl phosphate. The p81 and p32 were phosphorylated at histidine. The p44 had mainly phosphoserine and a small part of phosphohistidine. The phosphorylation profile was variable depending on the growth conditions. Furthermore, by a virulent phage Xp10 infection the quantity of phosphorylation increased: for phosphohistinine more than 10-fold, and for phosphoserine about 3-fold. Thus, in this bacterium phosphorylation may be linked with a physiological regulation system and with Xp10 phage development.     

The Status of Cassava Bacterial Blight Caused by Xanthomonas campestris pv. manihotis in Trinidad

Disease surveys conducted in Trinidad between 1985—1987 showed that Cassava Bacterial Blight (CBB) is present in all but one county of the country with disease severity ratings varying from 1—5 depending on day/night temperatures. Field and greenhouse screening identified varieties such as Point Fortin fine leaf and CMC 40 as being resistant whereas M col 22 was moderately resistant to susceptible. Using a combination of antiserum produced to whole cells of Xanthomonas campestris pv. manibotis and a broth enrichment technique, dissemination of the pathogen by flood water was confirmed. The pathogen was detected at distances of up to 300 meters from infected fields. The significance of this mode of pathogen dissemination in initiating primary infection in Trinidad is discussed.     

体细胞突变体HX-3抗水稻白叶枯病基因的鉴定

以感病杂交稻恢复系明恢63的成熟胚为外植体,利用离体筛选技术获得了抗水稻白叶枯病细胞突变体HX-3。连续8年以我国长江流域白叶枯病代表菌析浙173（IV型）对HX-3的R1到R9代进行抗性鉴定,HX-3的抗病性可以稳定遗传。抗性遗传分析表明HX-3的抗性由1对显性核基因控制。1999～2000年连续两年利用我国、菲律宾和日本的32个水稻白叶枯病菌株,测定HX-3及IRBB1等13个具不同显性抗病基因的近等基因系抗性,HX-3抗谱广,且与已知显性抗病基因的抗谱不同。在此基础上,以抗白叶枯病近等基因系IRBB4、IRBB7、CBB12和IRBB21和HX-3杂交,进行等位性分析,4个杂交组合的F2代均出现抗、感分离,说明HX-3与这4个基因不等位。综合以上研究结果,HX-3具有1个新的抗白叶枯病基因,暂命名为Xa-25(t）。     

High-quality mutant libraries of Xanthomonas oryzae pv. oryzae and X. campestris pv. campestris generated by an efficient transposon mutagenesis system

A novel transposon mutagenesis system for the phytopathogenic bacteria Xanthomonas oryzae pv. oryzae (Xoo) and X. campestris pv. campestris (Xcc) was developed using a Tn5-based transposome. A highly efficient transformation up to 10(6) transformants per microg transposon DNA was obtained. Southern blot and thermal asymmetric interlaced polymerase chain reaction analyses of Tn5 insertion sites suggested a random mode of transposition. The transposition was stable in the transformants for 20 subcultures. Eighteen thousand and 17000 transformants for Xoo and Xcc, respectively, were generated, corresponding to 4X ORF coverage of the genomes. The libraries will facilitate the identification of pathogenicity-related genes as well as functional genomic analysis in Xoo and Xcc.     

Characterization of Isolates of Bacterial Blight of Cotton (Xanthomonas campestris pv. malvacearum) from Nicaragua

Seeds from cotton plants infected with Xanthomonas campestris pv. malvacearum were collected in different parts of Nicaragua in 1986. When the seeds were homogenized the pathogen could not always be identified by dilution plating. Therefore, an enrichment of bacteria on the natural host was induced before isolation. The cotton seeds were shaken with water and sand for 2 days and then sown in sand for germination. Rather often the developing cotyledons showed typical water-soaked spots, from which the pathogen could be isolated easily. This new method needed more time but made it possible to detect a low level of bacterial infestation. Altogether, 42 bacterial isolates were obtained. For inoculation experiments suspensions with 5x10⁵ CFU - ml^?1 were infiltrated into cotton leaves. Incubations of inoculated plants in growth chambers, but not in greenhouses, resulted in typical and uniform disease symptoms (water-soaked leaf spots). Nine of the ten cotton differentials tested were highly susceptible to all the 42 bacterial isolates. Since only line 101-102B proved to be resistant, the Nicaraguan isolates of bacterial blight of cotton were characterized as race 18, of the pathogen. The main cotton cultivars grown in Nicaragua (H-373 and G-286) were strongly affected by the isolated bacterial strains. In order to reduce the disease incidence in Nicaragua, the cultivation of resistant cotton varieties is suggested.     

水稻白叶枯病生理生化机制的研究进展

以水稻-病原菌之间相互作用的两大方面寄主的侵染和宿主的防御作为文章线索,综述了近年来国内外有关水稻白叶枯病菌侵染机理与水稻防御机制的研究进展.     

Biological Role of Xanthomonadin Pigments in Xanthomonas campestris pv. Campestris

Previous studies have indicated that the yellow pigments (xanthomonadins) produced by phytopathogenic Xanthomonas bacteria are unimportant during pathogenesis but may be important for protection against photobiological damage. We used a Xanthomonas campestris pv. campestris parent strain, single-site transposon insertion mutant strains, and chromosomally restored mutant strains to define the biological role of xanthomonadins. Although xanthomonadin mutant strains were comparable to the parent strain for survival when exposed to UV light; after their exposure to the photosensitizer toluidine blue and visible light, survival was greatly reduced. Chromosomally restored mutant strains were completely restored for survival in these conditions. Likewise, epiphytic survival of a xanthomonadin mutant strain was greatly reduced in conditions of high light intensity, whereas a chromosomally restored mutant strain was comparable to the parent strain for epiphytic survival. These results are discussed with respect to previous results, and a model for epiphytic survival of X. campestris pv. campestris is presented.     

水稻类Tubby蛋白质在叶片生长和白叶枯病抗性反应中的表达

类Tubby蛋白质（Tubby-like protein,TLP）在动植物中广泛存在,暗示其在生命过程中发挥重要的作用。水稻（Oryza sativa）基因组中有14个TLP家族成员,首先制备了这些蛋白质的抗体,用免疫印迹方法检测了它们在水稻叶片不同生长时期的表达情况,揭示其表达模式;然后对Xa21介导的水稻白叶枯病抗性反应不同时间点进行检测,发现OsTLP2、OsTLP7、OsTLP8和OsTLP9等4个蛋白质的表达发生了变化;进一步比较它们在抗病、感病反应和对照处理中的表达情况,发现不同反应间的表达也有区别。该研究结果为阐释水稻TLP在叶片生长过程中的功能,尤其是在水稻-白叶枯病菌互作过程中的作用提供了重要线索。     

Abscisic Acid Promotes Susceptibility to the Rice Leaf Blight Pathogen Xanthomonas oryzae pv oryzae by Suppressing Salicylic Acid-Mediated Defenses

The plant hormone abscisic acid (ABA) is involved in a wide variety of plant processes, including the initiation of stress-adaptive responses to various environmental cues. Recently, ABA also emerged as a central factor in the regulation and integration of plant immune responses, although little is known about the underlying mechanisms. Aiming to advance our understanding of ABA-modulated disease resistance, we have analyzed the impact, dynamics and interrelationship of ABA and the classic defense hormone salicylic acid (SA) during progression of rice infection by the leaf blight pathogen Xanthomonas oryzae pv. oryzae (Xoo). Consistent with ABA negatively regulating resistance to Xoo, we found that exogenously administered ABA renders rice hypersusceptible to infection, whereas chemical and genetic disruption of ABA biosynthesis and signaling, respectively, led to enhanced Xoo resistance. In addition, we found successful Xoo infection to be associated with extensive reprogramming of ABA biosynthesis and response genes, suggesting that ABA functions as a virulence factor for Xoo. Interestingly, several lines of evidence indicate that this immune-suppressive effect of ABA is due at least in part to suppression of SA-mediated defenses that normally serve to limit pathogen growth. Resistance induced by the ABA biosynthesis inhibitor fluridone, however, appears to operate in a SA-independent manner and is likely due to induction of non-specific physiological stress. Collectively, our findings favor a scenario whereby virulent Xoo hijacks the rice ABA machinery to cause disease and highlight the importance of ABA and its crosstalk with SA in shaping the outcome of rice-Xoo interactions.     

Specific protein phosphorylation induced in Xanthomonas campestris pv. oryzae by bacteriophage Xp12

We have investigated the endogenous phosphorylation patterns of phosphorylated proteins of Xanthomonas campestris pv. oryzae induced by its bacteriophages. For bacteriophage Xp12-infected cells, at least three phosphoproteins with apparent molecular weights of 28, 28.5 and 45kDa were detected by in vitro labeling with [-³²P]-ATP. These Xp12-specific phosphoproteins only occurred with Xp12 infection, and were not shown in uninfected or Xp10-infected cells. The protein kinase(s) responsible could use either ATP or GTP as the nucleotide substrate with nearly the same efficiency. Magnesium was proved to be an essential factor for the phosphorylation. EGTA treatment excluding the possibility that the presumed protein kinase was calcium-dependent. Under our reaction conditions, the optimal phosphorylation occurred at pH 7 to 8, for 30 to 40 min at 25 to 37°C. The Xp12-specific protein phosphorylation hint the existence of a physiological regulation mechanism involved in the life cycle of bacteriophage Xp12. Furthermore, the presumed protein kinase was shown to be encoded by the genome of Xp12 rather than indirectly induced by Xp12 infection.     

A new protein subunit k for RNA polymerase from Xanthomonas campestris pv. oryzae

During the purification of RNA polymerase from Xanthomonas campestris pv. oryzae, a new subunit named k was found to be associated with this enzyme. The removal of subunit k from holoenzyme by DEAE-cellulose column chromatography results in a decrease in specific activity of the enzyme. The readdition of subunit k to subunit k-depleted holoenzyme results in restoration of enzymatic activity. Subunit k increase the activity of RNA polymerase; the activation was in proportion to the concentration of subunit k added. Antiserum against holoenzyme devoid of subunit k was prepared. This antiserum did not react with purified subunit k; therefore, subunit k may not be the proteolytic fragment of the beta, beta', sigma, or alpha subunit. When this antiserum was used to precipitate RNA polymerase obtained from a crude extract of bacterial cells, subunit k was coprecipitated as determined by sodium dodecyl sulfate gel electrophoretic analysis. The molecular mass of subunit k is approximately 29 kDa, and the molar ratio of beta:beta':sigma:alpha:k was estimated to be 1:1:1:2:4. When native Xp10 DNA was used as template, subunit k stimulated subunit k-depleted holoenzyme, but not core enzyme. When the synthetic polynucleotide poly[d(A-T)] was used, subunit k activated both subunit k-depleted holoenzyme and core enzyme. Subunit k also activated the binding of RNA polymerase to template DNA.