Transformation of the rice blast fungus Magnaporthe grisea to benomyl resistance

A transformation method based on a dominant selectable marker (benomyl resistance) was developed for the rice blast fungus Magnaporthe grisea. The heterologous gene for -tubulin from Neurospora crassa (pBT3) was used to obtain benomyl-resistant M. grisea transformants at a frequency of 20 to 30/g of DNA. Control transformations carried out with a plasmid conferring hygromycin resistance or a derivative of pBT3 containing a repetitive DNA sequence, yielded the same frequency of transformation as that of pBT3. Molecular analysis of the transformants indicated multiple integration of the vector DNA.     

Guanosine Diphosphate Mannose in Conformational Relation to Other Suger Nucleotides and Phosphates in Solution

Molecular conformational transition of GDPMan and solution conformation of α-d- mannopyranose moiety in Man-l-P and GDPMan were examined in relation to other sugar nucleotides and phosphates. GDPMan and other sugar nucleotides examined revealed changes in the optical rotation in sigmoidal curve in water by addition of urea. The change was reversible without significant decomposition and is attributable to dissociation of an ordered form into a random form. Optical conformational values in 8m urea solution were+116° for GDPMan, +58°~+79° for UDPGlc, +79° for UDPGal, +135°~+143° for UDPGlcNAc, and +138°~ +155° for UDPGIcA.

NMR analysis and periodate oxidation study revealed the ⁴C₁ conformation of α-d-hexopyranose moieties in Man-1-P, Glc-l-P, GDPMan, UDPGlcNAc and UDPGalNAc.     

The Nutritional Value of D-Amino Acid in the Chick Nutrition

The nutritional values of 16 D-amino acids in chick growth were studied on the purified diets containing crystalline amino acids as a sole source of nitrogen. Growth rate, feed consumption and nitrogen retention were measured. The nutritional values of D-amino acids were studied by comparing individually with the control groups fed on the diet containing all L-amino acids and negative control groups fed with the diet omitted the corresponding L-isomer. The following results were obtained. Essential amino acids: 1. Equal or almost equal nutritional value to the corresponding L-isomer; methionine, phenylalanine, leucine, proline. 2. Half nutritional value compared with L-isomer; valine. 3. Small nutritional value compared with L-isomer; tryptophan, isoleucine, histidine. 4. No nutritional value; lysine, threonine, arginine. Non-essential amino acid: 1. Equal or almost equal nutritional value to the corresponding L-isomer; serine, tyrosine, cystine. 2. There is a possibility that it has a slight growth retardation effect; alanine. 3. The growth retardation effect was found; aspartic acid.     

Establishment of a New Cross of the Rice Blast Fungus Derived from Japanese Differential Strain Ina168 and Hermaphroditic Rice Pathogen Guy11

Mating experiments between Magnaporthe grisea Japanese rice pathogens and Guy11, a hermaphroditic fertile rice pathogen, were done aimed at identification of avirulence genes. A cross named cross 2107 with thirty-six random progenies was obtained. Segregation analyses of genetic markers found that the cross was less suitable for genetic analysis. Backcrosses with cross 2107 progenies and Guy11 were done and another cross named cross 5307 with sixty-five progenies was obtained. A locus controlling kasugamycin resistance named Ksg1R was identified and used for a model case of genetic mapping. Bulked segregant analysis was done to find adjacent RAPD markers for mapping of the gene. Three adjacent markers to Ksg1R were obtained and a genetic map around the Ksg1R was made, but these markers were not located on a single chromosome. These results suggest that genetic analysis to identify a gene locus is available in cross 5307. Infection assay of parental strains of cross 5307 to Japanese differential rice cultivars suggested the possibility of genetic analysis of cultivar specificity toward four rice cultivars: Aichiasahi, Kusabue, Tsuyuake, and K59.     

稻瘟病菌致病突变体的REMI诱变与鉴定

以水稻“爱知旭”(Aichi—aShahi)为寄主,将带选择标记的质粒pCSN43和pBF101为外源DNA,利用限制酶诱导整合(REMI)这种新方法转化稻瘟病菌原生质体,从筛选到的数百个转化体中分离出3个与致病能力密切相关的突变体R2H65、R2H69和R281565。其中,R2H65和R2H69只产生畸形分生孢子,分生孢子的发育和附着胞的形成以及黑色素的合成均受到极大影响,致病性测试证明完全丧失致病能力;R281565的许多表型与野生型的相似,但致病能力却大大降低。     

靛红衍生物的合成及其对稻瘟菌的生物活性

以靛红为原料合成了系列3-亚胺基/亚甲基-吲哚-2-酮化合物及其Mannich碱衍生物,研究了它们在抗稻瘟菌方面的活性,发现了这两种类型的若干化合物有较好的抑制稻瘟菌孢子萌发的活性,初步讨论了构效关系。认为1位的羟甲基和胺甲基、3位的亚甲基是药效团,芳基亚甲基苯环上对位取代基、羟基取代基和吸电子取代基不利于活性的提高,邻位的供电子取代基有利于活性的提高。     

Allozyme variation in the endangered insular endemic Castilleja grisea

BACKGROUND AND AIMS: Genetic diversity in Castilleja grisea, an endangered, perennial herb endemic to San Clemente Island, California was investigated. Subsequent to the elimination of goats from the island in 1992, many populations of C. grisea have reappeared and have been increasing in size. METHODS: Nineteen populations were surveyed for their genotype at 19 allozyme loci. KEY RESULTS: At the taxon level, 57.9 % of loci are polymorphic with A(P) = 3.09 and H(E) = 0.137. Populations averaged 33.0 % polymorphic loci with A(P) = 2.43 and H(E) = 0.099. Most variation is found within rather than among populations (G(ST) = 0.128), although differentiation among populations is significant. Genetic identities range from I = 0.960 to I = 1.000 with mean I = 0.990. There is no significant relationship between genetic and geographic distance. Gene flow among populations is Nm = 2.50 based on private alleles and Nm = 1.70 based on F(ST). Outcrossing rates based on fixation indices average t = 1.01, indicating a primarily out-crossed mating system. CONCLUSIONS: The observed genetic variation is moderately high, unusually so for an insular endemic species, suggesting that C. grisea may not have lost substantial genetic variation during 150 years of overgrazing, and indicating that it is unlikely to be endangered by genetic factors.     

Tête à tête inside a plant cell: establishing compatibility between plants and biotrophic fungi and oomycetes

'Compatibility' describes the complementary relationship between a plant species and an adapted pathogen species that underlies susceptibility and which ultimately results in disease. Owing to elaborate surveillance systems and defence mechanisms on the plant side and a common lack of adaptation of many microbial pathogens, resistance is the rule and compatibility the exception for most plant-microbe combinations. While there has been major scientific interest in 'resistance' in the past decade, which has revealed many of its underlying molecular components, the analysis of 'compatibility', although intimately intertwined with 'resistance', has not been pursued with a similar intensity. Various recent studies, however, provide a first glimpse of the pivotal players and potential molecular mechanisms essential for compatibility in both the plant and parasite partners. In this review we highlight these findings with a particular emphasis on obligate biotrophic and hemibiotrophic fungal and oomycete pathogens and discuss novel strategies that might help to uncover further the molecular principles underlying compatibility to these highly specialized pathogens.     

A Novel Gene Involved in Lesion Formation in Magnaporthe grisea

To date, a number of genes that are expressed in the early stages of infection have been reported, while few genes that are expressed during the course of colonization, after the initiation of plant infection, have been studied. Plant inoculations, real‐time polymerase chain reaction (PCR), gene cloning, protein expression and bioinformatics analysis were combined to identify a novel gene, MgNIP04, in the rice blast fungus, Magnaporthe grisea. Bioinformatics analysis showed that the amino acid sequence contained a signal peptide. The wounded inoculation resulted in necrosis specks when the total expressed proteins including MBP‐MgNIP04 was inoculated on the wounded rice leaves, which demonstrated the protein directly interacted with rice. The real‐time PCR result of infected leaves showed that it is upregulated during late stages of infection of rice. Additionally, the copies of the gene expression absolute quantity of the gene were 7.61 × 10², 1.06 × 10³, 1.31 × 10³, 4.13 × 10³, and 4.00 × 10³, at 24, 48, 72, 96 and 168 h postinoculation (HPI), respectively. The higher copies of MgNIP04 were found at 96 and 168 HPI. The copies of MgNIP04 in mycelia of Y98‐63C, Y99‐16, 94‐64‐1b and 95‐23‐4a were significantly lower than those of infected leaves. Through the above bioinformatics and experimental analysis, our result suggested that the MgNIP04 was novel pathogenicity‐related protein in rice blast fungus.