Exploring Durable Genetic Resistance against Leaf Rust through Phenotypic Characterization and Lr34Linked STS Marker in Wheat Germplasm / Explorando a resistência genética duradoura contra a ferrugem da folha através da caracterização fenotípica e do STS marker relacionado ao (GENE) LR34 do germoplasma do trigo

Present study was aimed to screening the population of 25 wheat genotypes from Baluchistan region of Pakistan along with five commercial cultivars for leaf rust adult plant resistance (APR) through gene postulation using natural inoculation of Puccinia triticina Erikss local pathotype. Infection severity was recorded on scale in comparison with susceptible control "Morroco" cultivar. On the basis of phenotypic score, seven accessions and four varieties (Zardana-89, Sariab-92, Zarlashta-99 and Raskoh-05) with AUDPC values up to 20% were characterized as resistant genotypes. Coefficient of infection (CI) score ranged from 0-10 for some accessions and cultivars showing high level of adult plant resistance. Furthermore, bi-allelic STS marker csLV34 having close linkage with Lr34 (0.4cM). This marker amplified one gene specific allele of 150bp in 21 genotypes, including 19 accessions and two commercial varieties (Sariab-92 and Zarghoon-79) which confirmed presence of Lr34 gene conferring adult plant resistance against leaf rust. The rust pathogenicity scale varied for accessions from resistant to moderately susceptible. However, beside Lr34, phenotypic gene postulation, in combination with marker assisted selection for leaf rust resistance, has revealed presence of some other unknown resistance genes in local wheat germplasm which signified its use in wheat improvement programs both locally and abroad.

O presente estudo teve como objetivo a triagem da população de 25 genótipos de trigo do Baluchistão, região do Paquistão, juntamente com cinco cultivares comerciais para o estudo da resistência à ferrugem da folha em plantas adultas (leaf rust adult plant resistance, APR, em inglês) através da postulação gênica usando a inoculação natural do patótipo local da Puccinia triticina Erikks. A gravidade da infecção foi registrada na escala em comparação ao cultivar de controle suscetível "Morroco". Com base na pontuação fenotípica, sete acessões e quatro variedades (Zardana-89, Sariab-92, Zarlashta-99 and Raskoh-05) com valores de AUDPC (area under the disease progress curve, em inglês) até 20% foram caracterizados como genótipos resistentes. A pontuação do coeficiente de infecção (CI) variou no intervalo de 0-10 para algumas acessões e cultivares evidenciando uma elevada resistência nas plantas adultas. Além disso, o STS marker para o csLV34 bi-alélico demonstrou uma ligação estreita com o Lr34 (0.4cM). Este marcador amplificou um alelo específico do gene do 150bp em 21 genótipos, incluindo 19 acessões e duas variedades comerciais (Sariab-92 and Zarghoon-79) o que confirmou a presença do gene Lr34 conferindo resistência às plantas adultas contra a ferrugem da folha. A escala de patogenicidade da ferrugem para as acessões de resistente a moderadamente suscetível. Contudo, além do Lr34, a postulação gênica fenotípica, em combinação com a seleção auxiliada (ou assistida) por marcadores para a resistência da ferrugem da folha, revelou a presença de outros genes resistentes desconhecidos no germoplasma do trigo local o que justifica a sua utilização em programas de melhoramento do trigo tanto a nível local quanto a nível internacional.

Identification and characterization of rhizospheric microbial diversity by 16S ribosomal RNA gene sequencing

In the present study, samples of rhizosphere and root nodules were collected from different areas of Pakistan to isolate plant growth promoting rhizobacteria. Identification of bacterial isolates was made by 16S rRNA gene sequence analysis and taxonomical confirmation on EzTaxon Server. The identified bacterial strains were belonged to 5 genera i.e. Ensifer, Bacillus, Pseudomona, Leclercia and Rhizobium. Phylogenetic analysis inferred from 16S rRNA gene sequences showed the evolutionary relationship of bacterial strains with the respective genera. Based on phylogenetic analysis, some candidate novel species were also identified. The bacterial strains were also characterized for morphological, physiological, biochemical tests and glucose dehydrogenase (gdh) gene that involved in the phosphate solublization using cofactor pyrroloquinolone quinone (PQQ). Seven rhizoshperic and 3 root nodulating stains are positive for gdh gene. Furthermore, this study confirms a novel association between microbes and their hosts like field grown crops, leguminous and non-leguminous plants. It was concluded that a diverse group of bacterial population exist in the rhizosphere and root nodules that might be useful in evaluating the mechanisms behind plant microbial interactions and strains QAU-63 and QAU-68 have sequence similarity of 97 and 95% which might be declared as novel after further taxonomic characterization.

Bioinformatics based structural characterization of glucose dehydrogenase (gdh) gene and growth promoting activity of Leclercia sp. QAU-66

Glucose dehydrogenase (GDH; EC 1.1. 5.2) is the member of quinoproteins group that use the redox cofactor pyrroloquinoline quinoine, calcium ions and glucose as substrate for its activity. In present study, Leclercia sp. QAU-66, isolated from rhizosphere of Vigna mungo, was characterized for phosphate solubilization and the role of GDH in plant growth promotion of Phaseolus vulgaris. The strain QAU-66 had ability to solubilize phosphorus and significantly (p < 0.05) promoted the shoot and root lengths of Phaseolus vulgaris. The structural determination of GDH protein was carried out using bioinformatics tools like Pfam, InterProScan, I-TASSER and COFACTOR. These tools predicted the structural based functional homology of pyrroloquinoline quinone domains in GDH. GDH of Leclercia sp. QAU-66 is one of the main factor that involved in plant growth promotion and provides a solid background for further research in plant growth promoting activities.