rbcS SRS4 promoter from Glycine max and its expression activity in transgenic tobacco

Cui, Xiyan; Chen, Z Y; Wu, Lintao; Liu, X Q; Dong, Yan; Wang, F W; Li, H Y

doi:10.4238/2015.july.3.15

Cited by 5 publications

(4 citation statements)

References 31 publications

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“…Green tissue specific promoters direct the expression preferentially in shoots and limit the expression in roots, where TI production is not needed. A previous study also showed similar gene expression patterns in leaves comparing 35S and rbcS-SRS4 promoter (Cui et al, 2015).…”

Section: Discussionsupporting

confidence: 63%

“…Each soybean TI gene was also cloned under the control of a green tissue-specific promoter. Soybean green-tissue specific promoter (1775 bp) of ribulose-1,5bisphosphate carboxylase small subunit gene SRS4 (rbcs-SRS4) (Cui et al, 2015) was amplified by PCR from soybean cv. 'Williams 82' using primers incorporating specific restriction enzymes (SacI and SpeI) (Supplementary Table S1).…”

Section: Expression Vector Constructionsmentioning

confidence: 99%

See 1 more Smart Citation

Overexpression of soybean trypsin inhibitor genes decreases defoliation by corn earworm (Helicoverpa zea) in soybean (Glycine max) and Arabidopsis thaliana

Sultana

Mazarei²,

Jurat-Fuentes³

et al. 2023

Front. Plant Sci.

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Trypsin inhibitors (TIs) are widely distributed in plants and are known to play a protective role against herbivores. TIs reduce the biological activity of trypsin, an enzyme involved in the breakdown of many different proteins, by inhibiting the activation and catalytic reactions of proteins. Soybean (Glycine max) contains two major TI classes: Kunitz trypsin inhibitor (KTI) and Bowman-Birk inhibitor (BBI). Both genes encoding TI inactivate trypsin and chymotrypsin enzymes, which are the main digestive enzymes in the gut fluids of Lepidopteran larvae feeding on soybean. In this study, the possible role of soybean TIs in plant defense against insects and nematodes was investigated. A total of six TIs were tested, including three known soybean trypsin inhibitors (KTI1, KTI2 and KTI3) and three genes encoding novel inhibitors identified in soybean (KTI5, KTI7, and BBI5). Their functional role was further examined by overexpression of the individual TI genes in soybean and Arabidopsis. The endogenous expression patterns of these TI genes varied among soybean tissues, including leaf, stem, seed, and root. In vitro enzyme inhibitory assays showed significant increase in trypsin and chymotrypsin inhibitory activities in both transgenic soybean and Arabidopsis. Detached leaf-punch feeding bioassays detected significant reduction in corn earworm (Helicoverpa zea) larval weight when larvae fed on transgenic soybean and Arabidopsis lines, with the greatest reduction observed in KTI7 and BBI5 overexpressing lines. Whole soybean plant greenhouse feeding bioassays with H. zea on KTI7 and BBI5 overexpressing lines resulted in significantly reduced leaf defoliation compared to non-transgenic plants. However, bioassays of KTI7 and BBI5 overexpressing lines with soybean cyst nematode (SCN, Heterodera glycines) showed no differences in SCN female index between transgenic and non-transgenic control plants. There were no significant differences in growth and productivity between transgenic and non-transgenic plants grown in the absence of herbivores to full maturity under greenhouse conditions. The present study provides further insight into the potential applications of TI genes for insect resistance improvement in plants.

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Section: Discussionsupporting

confidence: 63%

Section: Expression Vector Constructionsmentioning

confidence: 99%

Overexpression of soybean trypsin inhibitor genes decreases defoliation by corn earworm (Helicoverpa zea) in soybean (Glycine max) and Arabidopsis thaliana

Sultana

Mazarei²,

Jurat-Fuentes³

et al. 2023

Front. Plant Sci.

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“…The soybean cultivar MON 87701-2 was produced by transfering the cry1Ac gene using a gene promoter rbcS4 control of the species Arabidopsis thaliana. This promoter expresses the characteristics of the gene of interest at high levels in green tissue (Cui et al 2015). The protein sequence is 99% identical to the Cry1Ac protein from B. thuringiensis kurstaki.…”

Section: Bt Soybeanmentioning

confidence: 93%

Genetically modified soybean expressing insecticidal protein (Cry1Ac): Management risk and perspectives

Martins-Salles

Machado

Massochin-Pinto

et al. 2017

FACETS

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A Bt soybean has been recently developed, thus, efficiently regulating the populations of major lepidopteran pests. However, in other cases, these benefits have been reduced or lost because of the rapid evolution of pest resistance to the Bt toxins in transgenic crops. When pest populations are exposed to Bt crops and to refuges (non-Bt plants), the evolution of resistance is governed by the fitness of resistant individuals relative to susceptible individuals for both the presence and absence of Bt toxins. One major ecological concern regarding the biosafety of Bt crops on the environment is their potential effects on non-target organisms, especially predators and parasitoids that play an important role in pest control. This information is important for supporting insect resistance management (IRM) programs and for improving agricultural practices in a crop production system with Bt plants. Before the use of Bt plants for insect pest control in Brazil is adopted, IRM programs should be established to ensure the sustainability of this technology for integrated pest management (IPM). This review presents data on Bt soybean and lepidopteran pests as well as on the importance of natural enemies as a form of biological control, and applications for IPM and IRM.

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“…In this experiment, two Bt genes were constructed on a vector by utilizing the matrix attachment region (MAR) structure, applying different promoters (Cui et al, 2015) and enhancer sequences, and changing the open reading frame (ORF) sequence of exogenous genes. Four different types of vectors were constructed and transferred to tobacco, to obtain transgenic plants.…”

Section: Introductionmentioning

confidence: 99%

Effect of dual Bt-expression transformation vectors on transgene expression in tobacco

et al. 2016

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ABSTRACT. This study aimed to determine the influence of vector structure on dual Bt gene expression and establish an efficient expression vector using Cry1Ac and Cry3A genes. Four vectors (N4, N5, N10, and S23) were developed and used for genetic transformation of tobacco to obtain insect-resistant transgenic lines. The vectors were constructed using the MAR structure, applying different promoter and enhancer sequences, and changing the transgene open-reading frame sequence. The average Cry1Ac toxalbumin expression quantity was 67 times higher in N5 than in N4 transgenic lines (8.77 and 0.13 µg/g, respectively). In contrast, the average Cry3A toxalbumin expression quantity was 1.5 times higher in N4 than in N5 lines (12.70 and 8.21 µg/g, respectively). The sequences of both Bt genes significantly influenced toxalbumin expression, although upstream Bt genes presented lower expression levels. The average Cry1Ac toxalbumin content was 13 times higher in the transgenic lines of AtADH 5'-nontranslated sequence N5 (8.77 mg/g) than in the omega N10 lines (0.67 mg/g). Furthermore, the average Cry1Ac toxalbumin content was 5 times higher in MAR N5 than in non-MAR S23 lines (8.77 and 1.63 mg/g, respectively). The average Cry3A toxalbumin content was 1.3 times higher in N5 than in S23 lines (8.21 and 6.48 mg/g, respectively). Moreover, toxalbumin expression levels differed significantly among the S23-transformed lines. The MAR structure applied on both ends of the genes increased both the level and stability of exogenous gene expression. In conclusion, N5 was the most optimal of the four tested vectors.

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rbcS SRS4 promoter from Glycine max and its expression activity in transgenic tobacco

Cited by 5 publications

References 31 publications

Overexpression of soybean trypsin inhibitor genes decreases defoliation by corn earworm (Helicoverpa zea) in soybean (Glycine max) and Arabidopsis thaliana

Overexpression of soybean trypsin inhibitor genes decreases defoliation by corn earworm (Helicoverpa zea) in soybean (Glycine max) and Arabidopsis thaliana

Genetically modified soybean expressing insecticidal protein (Cry1Ac): Management risk and perspectives

Effect of dual Bt-expression transformation vectors on transgene expression in tobacco

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