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Population genetic structure of Chinese Puccinia triticina races based on multi-locus sequences
LIU Tai-guo, GE Run-jing, MA Yu-tong, LIU Bo, GAO Li, CHEN Wan-quan
2018, 17 (08): 1779-1789.   DOI: 10.1016/S2095-3119(18)61923-9
Abstract313)      PDF in ScienceDirect      
Puccinia triticina, the causal agent of wheat leaf rust, is one of the most devastating rust fungi attacking wheat worldwide.  Seventy-six isolates of the wheat leaf rust pathogen from Yunnan, Sichuan, Gansu and Henan provinces, China, were tested on wheat leaf rust differentials and the population structure was analyzed using four presumably neutral partial sequence markers such as elongation factor-1α (EF-1α), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), β-tubulin (TUB) and the second largest RNA polymerase subunit (RPB2).  The phenotypic diversity of Yunnan and Sichuan populations was higher than that of Gansu and Henan populations.  The four populations were separated into two clusters based on the pathogenic data.  A total of 12 single nucleotide polymorphisms (SNPs) and 32 haplotypes were identified among the four sequences.  The 32 haplotypes were divided into two clusters in a neighbor-joining tree.  Bayesian analyses also identified two clusters.  Pairwise Fst between populations in different regions were significantly different (P<0.05).  Analysis of molecular variance (AMOVA) indicated that 68% of the total genetic variation was within populations. 
 
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Postulation of seedling leaf rust resistance genes in 84 Chinese winter wheat cultivars
REN Xiao-li, LIU Tai-guo, LIU Bo, GAO Li, CHEN Wan-quan
2015, 14 (10): 1992-2001.   DOI: 10.1016/S2095-3119(14)61002-9
Abstract1707)      PDF in ScienceDirect      
Wheat leaf rust (caused by Puccinia triticina) is one of the most important fungal diseases in China. There are tens of winter wheat cultivars which are approved to be released by the government at a national level and more than 100 wheat cultivars at the provincial level. But there is no information about leaf rust (Lr) genes in these cultivars, which makes it difficult for farmers and breeders to select which cultivars they should plant in their fields and use in their breeding programs. The objective of this paper was to identify the leaf rust resistant genes at seedling stage present in the 84 commercial wheat cultivars from China that have been released in the past few years. A set of 20 near isogenic lines with Thatcher background and 6 lines with known Lr genes were used to test the virulence of 12 races of P. triticina (Pt). By comparing the infection types (ITs) produced on the 84 cultivars by the 12 Pt races with the ITs on the differential sets, the Lr genes were postulated. In addition, 8 molecular markers of Lr genes such as Lr9, Lr10, Lr19, Lr20, Lr21, Lr24, Lr26 and Lr29, which are closely linked to or co-segregated with the Lr gene, were used for further validation of the genes in the 84 Chinese winter wheat cultivars. Twelve Lr genes, including Lr1, Lr3, (Lr3bg), (Lr3ka), Lr11, Lr13, Lr14a, Lr16, Lr26, Lr27, Lr30 and Lr31 were postulated to be present either singly or in combinations in these Chinese wheat cultivars. Lr3 and Lr26 were detected most often in the tested cultivars, with frequencies of 51.2 and 38.1%, respectively. No wheat Lr genes were detected in 16 cultivars, and 4 cultivars may carry unknown Lr genes other than those used in this study. Lr9, Lr20, Lr21, Lr24, Lr25 and Lr29 were not present in any of the 84 tested accessions.
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A FIASCO-Based Approach for Detection and Diagnosis of Puccinia graminis f. sp. tritici in China
LIU Tai-guo, WANG Xi, GAO Li, LIU Bo, CHEN Wan-quan , XIANG Wen-sheng
2014, 13 (11): 2438-2444.   DOI: 10.1016/S2095-3119(14)60895-9
Abstract1573)      PDF in ScienceDirect      
Stem or black rust of wheat, caused by the fungus Puccinia graminis Pers. f. sp. tritici Eriks. & E. Henn. (Pgt), has historically caused severe losses to wheat (Triticum aestivum L.) production worldwide. In the Fujian and Guangdong provinces of China, six moderate-to-severe epidemics of wheat stem rust have occurred, which caused destructive losses of wheat between 1949 and 1966, although these were brought under control by integrated management. A rapid and reliable detection of the pathogen will contribute to the accurate forecast and seasonal control of this disease. The objective of this study was to develop a diagnostic molecular marker generated from simple sequence repeats (SSR) for the early rapid identification of P. graminis. The genomic DNA of P. graminis, Puccinia striiformis, Puccinia triticina and seven other species was amplified by a pair of SSR primers generated by the FIASCO (fast isolation by AFLP sequences containing repeats) enrichment protocol. The primer set Pgtw (f)/ Pgtw (r) generated a polymorphic pattern displaying a 330-bp DNA fragment specific for P. graminis whereas no DNA fragment was obtained from other non-target wheat fungal pathogens. The detection limit of the primer was 1 ng DNA in a 25-mL PCR reaction. The SSR markers of P. graminis can also be used to detect the presence of latent hyphae in Pgt-infected wheat leaves as early as 30 h post-inoculation. A rapid approach to distinguish P. graminis from similar pathogenic fungi would be anticipated in further study.
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Jellyfish Green Fluorescent Protein (GFP) as a Reporter for Fusarium gramminearum Development on Wheat
QI Jun-xian, LIU Tai-guo, XU Ying, CHEN Huai-gu, GAO Li, LIU Bo , CHEN Wan-quan
2014, 13 (10): 2177-2183.   DOI: 10.1016/S2095-3119(14)60875-3
Abstract1128)      PDF in ScienceDirect      
The plasmid pGPDGFP under the control of pgpdA promotor was used together with vector pAN7-1 containing the hygromycin resistance cassette to co-transform protoplasts of HG1, Fusarium graminearum from Hubei Province, China. Twelve out of 14 hygromycin-resistant transformants showed green signal under the UV light and contained one or several copies of gfp, as indicated by Southern analysis of genomic DNA digested with different restriction enzymes and hybridized to the gfp probe. A single gfp copy transformant (HG1C5) was selected for further evaluation of 80 Chinese wheat cultivars or advanced lines. The results showed different resistance type to F. graminearum were observed. GFP signals observed in the rachis and adjacent spikes of 70 Chinese wheat lines such as Chuanchongzu 104 indicated both type I (host resistance to the initial infection by the fungus) and type II (resistance to the spread of FHB symptoms within an infected spike) were not observed. While other 10 lines showed type II resistance to F. graminearum with GFP signals only in inoculated spikelets. Development of the mycelium can be intuitively observed and the resistance of wheat to F. graminearum can be identified at 7 days post inoculation (dpi) in this way. The results showed no differences were evaluated between the transformed HG1C5 and the non-transgene artificial inoculation by SAS paired chi-square test and McNemar’s test (P=0.0625).
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