Optimization of the Cry2Aa gene and development of insect-resistant and herbicide-tolerant photoperiod-sensitive genic male sterile rice

Abstract

Weng L., Deng L., Lia F., Xiao G. (2014): Optimization of the Cry2Aa gene and development of insect-resistant and herbicide-tolerant photoperiod-sensitive genic male sterile rice. Czech J. Genet. Plant Breed., 50: 19-25.In this study, an optimized Cry2Aa gene was obtained after codon optimization based on the preferred codons in rice. The novel fusion gene Cry2Aa # was designed by adding the sequence coding the signal peptide of PR1a at the 5' end and the endoplasmic reticulum retention signal peptide KDEL at the 3' end of the optimized Cry2Aa gene, respectively. The Cry2Aa # and Bar genes were transformed into 4008S, a photoperiod-sensitive genic male sterile (PGMS) line in rice, using the Agrobacterium transfer method. A total of 65 regenerated plantlets confirmed by PCR were produced, in which eight transgenic lines had single-copy insertions as confirmed by Southern blot analysis. The high variability of Cry2Aa # gene expression was observed among independent transgenic lines with single-copy insertion, and the spatiotemporal difference of Cry2Aa protein expression was discovered in each transgenic line. The results showed that the transgenic lines were highly resistant to glufosinate, rice leaf roller and striped stem borer, which not only confirmed the effective optimization of the Cry2Aa gene but also produced a useful germplasm for breeding insect-resistant and herbicide-tolerant hybrid rice varieties.

Transgenic rice expressing a novel phytase-lactoferricin fusion gene to improve phosphorus availability and antibacterial activity

“…The PAT protein contents in the leaves of the glufosinate resistant plants in T 1 generation at tillering stage were between 50.45-69.05 μg g -1 , with an average content of 62.87 μg g -1 , which was much higher than the average content of CK-BPL9K (0.03 μg g -1 ), and higher than that of the reported transgenic rice B2A68 with 41.05 μg g -1 PAT protein in its leaves (Fig. 5-B) (Weng et al 2014); meanwhile the weaker resistant plant BP9K-2 got a PAT protein content (0.68 μg g -1 ) just a little bit higher than that of CK-BP9K (0.05 μg g -1 ). Results of going-on analysis of the segregated populations of T 2 generation from BPL9K-3 and BP9K-1 showed that: The PAT protein contents of leaves from BPL9K-3-1-BPL9K-3-7 were between 78.52-92.78 μg g -1 , with the average content of 85.09 μg g -1 ; the PAT protein contents of leaves from BP9K-1-1-BP9K-1-7 were between 72.60-93.39 μg g -1 , with the average content of 82.83 μg g -1 ; the PAT protein contents in leaves of segregants without foreign genes from the T 2 generation of BPL9K-3 and BP9K-1 were extremely low, with an average content of 0.05 μg g -1 (Fig.…”

Transient Expression of Plant-Codon-Optimized Cry2ab39 Gene by Agroinfiltration in N. Benthamiana

“…The A-T percentage of wild-type Cry genes is much higher than in a typical plant gene. These A-T-rich regions in bacterial genes might resemble plant introns or potential plant polyadenylation signal sequences and therefore cause premature termination of translation and subsequent degradation of the target mRNA [24,[35][36][37] . In addition, different preferences in codon usage of plant and bacterial systems lead to ineffective translation [35,37] .…”

Produção de plantas transgênicas de citros expressando toxina de <i>Bacillus thuringiensis</i> visando ao controle de <i>Diaphorina citri</i>, inseto vetor do agente causal do <i>huanglongbing</i> (HLB)

“…O processo como um todo envolve diversos eventos que devem ser concluídos várias horas após a ingestão, a fim de levar à morte do inseto (JISHA; SMITHA; BENJAMIN, 2013). Sabendo dessa diferença de expressão de genes após a otimização de códons, vários pesquisadores têm utilizado a otimização de códons de genes de Bt visando à aplicação na transgenia de plantas de diversas culturas (JABEEN et al, 2010;WENG et al, 2014;SINGH et al, 2016;DAS et al, 2017), pois a mesma é de grande relevância.…”