Beet cyst nematode (Heterodera schachtii Schm.) is one of the most damaging pests in sugar beet growing areas around the world. The Hs1pro-1 and cZR3 genes confer resistance to the beet cyst nematode, and both were cloned from sugar beet translocation line (A906001). The translocation line carried the locus from B. procumbens chromosome 1 including Hs1pro-1 gene and resistance gene analogs (RGA), which confer resistance to Heterodera schachtii. In this research, BvHs1pro-1 and BvcZR3 genes were transferred into oilseed rape to obtain different transgenic lines by A. tumefaciens mediated transformation method. The cZR3Hs1pro-1 gene was pyramided into the same plants by crossing homozygous cZR3 and Hs1pro-1 plants to identify the function and interaction of cZR3 and Hs1pro-1 genes. In vitro and in vivo cyst nematode resistance tests showed that cZR3 and Hs1pro-1 plants could be infested by beet cyst nematode (BCN) juveniles, however a large fraction of penetrated nematode juveniles was not able to develop normally and stagnated in roots of transgenic plants, consequently resulting in a significant reduction in the number of developed nematode females. A higher efficiency in inhibition of nematode females was observed in plants expressing pyramiding genes than in those only expressing a single gene. Molecular analysis demonstrated that BvHs1pro-1 and BvcZR3 gene expressions in oilseed rape constitutively activated transcription of plant-defense related genes such as NPR1 (non-expresser of PR1), SGT1b (enhanced disease resistance 1) and RAR1 (suppressor of the G2 allele of skp1). Transcript of NPR1 gene in transgenic cZR3 and Hs1pro-1 plants were slightly up-regulated, while its expression was considerably enhanced in cZR3Hs1pro-1 gene pyramiding plants. The expression of EDS1 gene did not change significantly among transgenic cZR3, Hs1pro-1 and cZR3Hs1pro-1 gene pyramiding plants and wild type. The expression of SGT1b gene was slightly up-regulated in transgenic cZR3 and Hs1pro-1 plants compared with the wild type, however, its expression was not changed in cZR3Hs1pro-1 gene pyramiding plant and had no interaction effect. RAR1 gene expression was significantly up-regulated in transgenic cZR3 and cZR3Hs1pro-1 genes pyramiding plants, but almost no expression was found in Hs1pro-1 transgenic plants. These results show that nematode resistance genes from sugar beet were functional in oilseed rape and conferred BCN resistance by activation of a CC-NBS-LRR R gene mediated resistance response. The gene pyramiding had enhanced resistance, thus offering a novel approach for the BCN control by preventing the propagation of BCN in oilseed rape. The transgenic oilseed rape could be used as a trap crop to offer an alternative method for beet cyst nematode control.
Drought stress is a common issue that affects agriculture production. The soybean plant is one of the drought-sensitive crops that could lose the yield of up to 40% under severe drought years. Improvements in drought tolerance have been one of the main objectives of breeding programs in soybean. In this study, 62 soybeans [Glycine max (L.) Merri.] germplasms including landraces and elite cultivars were evaluated by analyzing canopy and root morphological characteristics at seedling stage under polyethylene glycol (2.5 M PEG 8000, − 0.54 MPa) simulated drought stress. The results showed that the wilting index of sixty-two soybean germplasms showed significant polymorphism differences after 4 days PEG treatment. The lowest wilting index was 2.75 in Tiefeng 31, while the largest reached 9 in Fengdou 93 and Songzidou. The wilting index was positively correlated with plant height, electrolyte leakage, and negatively correlated with a dry weight of the above-ground part, stomatal conductance, and transpiration rate by regression analysis. The root system architecture (RSA) study showed root-canopy ratio, root length, number of the lateral roots, root surface area, and root volume were significantly negatively correlated with the wilting index and positively correlated with the root volume. Based on these results, sixty-two soybean germplasms were comprehensively evaluated for drought-tolerant cultivar by the principal component analysis (PCA). The results showed that Tiefeng 31 was the most drought-tolerant elite cultivar. However, Fengdou 93 was the most droughtsensitive elite variety. The drought tolerance screening results were consistent with the wilting index and RSA analysis, especially the root length. Both droughts tolerant and sensitive elite cultivar could be further used to breed drought-tolerant germplasms and to clarify the drought tolerance mechanism in soybean.
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