ABSTRACT
The soybean ubiquitous urease (encoded by GmEu4) is responsible for recycling metabolically derived urea. Additional biological roles have been demonstrated for plant ureases, notably in toxicity to other organisms. However, urease enzymatic activity is not related to its toxicity. The role of GmEu4 in soybean susceptibility to fungi was investigated in this study. A differential expression pattern of GmEu4 was observed in susceptible and resistant genotypes of soybeans over the course of a Phakopsora pachyrhizi infection, especially 24 h after infection. Twenty-nine adult, transgenic soybean plants, representing six independently transformed lines, were obtained. Although the initial aim of this study was to overexpress GmEu4, the transgenic plants exhibited GmEu4 co-suppression and decreased ureolytic activity. The growth of Rhizoctonia solani, Phomopsis sp., and Penicillium herguei in media containing a crude protein extract from either transgenic or non-transgenic leaves was evaluated. The fungal growth was higher in the protein extracts from transgenic urease-deprived plants than in extracts from non-transgenic controls. When infected by P. pachyrhizi uredospores, detached leaves of urease-deprived plants developed a significantly higher number of lesions, pustules and erupted pustules than leaves of non-transgenic plants containing normal levels of the enzyme. The results of the present work show that the soybean plants were more susceptible to fungi in the absence of urease. It was not possible to overexpress active GmEu4. For future work, overexpression of urease fungitoxic peptides could be attempted as an alternative approach.
Subject(s)
Basidiomycota/growth & development , Glycine max/enzymology , Plant Diseases/microbiology , Urease/metabolism , Biological Assay , DNA, Bacterial/genetics , Disease Resistance/genetics , Gene Expression Regulation, Plant , Genetic Vectors/genetics , Plant Diseases/genetics , Plant Leaves/microbiology , Plant Proteins/genetics , Plant Proteins/metabolism , Plants, Genetically Modified , Recombination, Genetic/genetics , Glycine max/genetics , Glycine max/microbiology , Transformation, Genetic , Transgenes/genetics , Urea/metabolismABSTRACT
In a greenhouse experiment, morpho-anatomical and micromorphometrical analyses of two soybean cultivars, MG/BR46 (Conquista) and BR16-tolerant and sensitive to drought, respectively-were used to study their water-deficit-tolerance strategies. Drought treatments were applied at reproductive stages from R2 to R7, where evaluations were conducted at 30 days and 45 days after stress started, respectively. The total length of Conquista plants (shoot + root) was greater than of BR16 plants. Pod dry weight was adversely affected due to the lack of moisture, decreasing productivity even of Conquista plants. Both the cultivars had normal development of root hairs; however, it was observed a decrease in the cortex:central cylinder ratio in BR16 stressed for 30 days, and they also showed similar leaflet thickness and stomata distribution. Differences in drought tolerance observed between the two cultivars seemed to be related to factors other than morphological traits since this species has a short lifecycle.
Análises morfo-anatômicas e micromorfométricas de duas cultivares de soja, MG/BR46 (Conquista) e BR16tolerante e sensível à seca, respectivamente, em experimento conduzido em casa de vegetaçãoforam feitas para estudar as diferentes estratégias de tolerância ao déficit hídrico. Tratamentos de seca foram aplicados no estádio reprodutivo R2 e R7, onde avaliações foram conduzidas em 30 dias e 45 dias após o inicio do estresse, respectivamente. O comprimento total das plantas da cultivar Conquista (parte aérea e raiz) foi maior do que das plantas da cultivar BR16. A massa seca da vagem foi adversamente afetada pelo déficit hídrico, diminuindo a produtividade das plantas da cultivar Conquista. Ambas as cultivares tiveram o desenvolvimento de pêlos radiciais normais e, uma diminuição da razão córtex:cilindro central foi observada em BR16 estressada por 30 dias, além de apresentar uma espessura do folíolo e distribuição dos estômatos normais. Diferenças na tolerância à seca observada entre as duas cultivares devem estar relacionadas também a outros fatores, alm das características morfológicas, já que esta espécie possui um ciclo de vida curto.
ABSTRACT
Drought cause serious yield losses in soybean (Glycine max), roots being the first plant organ to detect the water-stress signals triggering defense mechanisms. We used two drought induction systems to identify genes differentially expressed in the roots of the drought-tolerant soybean cultivar MG/BR46 (Conquista) and characterize their expression levels during water deficit. Soybean plants grown in nutrient solution hydroponically and in sand-pots were submitted to water stress and gene expression analysis was conducted using the differential display (DD) and real time polymerase chain reaction (PCR) techniques. Three differentially expressed mRNA transcripts showed homology to the Antirrhinum majus basic helix-loop-helix transcription factor bHLH, the Arabidopsis thaliana phosphatidylinositol transfer protein PITP and the auxin-independent growth regulator 1 (axi 1). The hydroponic experiments showed that after 100 min outside the nutrient solution photosynthesis completely stopped, stomata closed and leaf temperature rose. Both stress induction treatments produced significant decrease in the mitotic indices of root cells. Axi 1, PITP and bHLH were not only differentially expressed during dehydration in the hydroponics experiments but also during induced drought in the pot experiments. Although, there were differences between the two sets of experiments in the time at which up or down regulation occurred, the expression pattern of all three transcripts was related. Similar gene expression and cytological analysis results occurred in both systems, suggesting that hydroponics could be used to simulate drought detection by roots growing in soil and thus facilitate rapid and easy root sampling.