High resistance of transgenic cabbage plants with a synthetic <i>cry1Ia8</i>
 gene from <i>Bacillus thuringiensis</i>
 against two lepidopteran species under field conditions

Yi, Dengxia; Yang, Weijie; Tang, Jun; Wang, Li; Fang, Zhiyuan; Liu, Yumei; Zhuang, Mu; Zhang, Yangyong; Yang, Limei

doi:10.1002/ps.3994

Cited by 7 publications

(3 citation statements)

References 19 publications

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“…For most cruciferous crops, a complete genetic transformation system has been established; However, for a few crop species, these systems have not yet been perfected. Agrobacterium-mediated transformation and CRISPR/ Cas9 gene editing have been widely applies to rape (Probsting et al, 2020), cabbage (cry1Ia8, cry1C) (Yi et al, 2016;, and broccoli (Cry1Ac) (Cao et al, 2002). However, research on transformation system of Chinese cabbage is lagging behind the other Brassica crops, such as rapeseeds, cabbage and so on, so VIGS must be used to identify functional genes (Murata and Orton, 1987).…”

Section: Introductionmentioning

confidence: 99%

A highly efficient genetic transformation system for broccoli and subcellular localization

et al. 2023

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IntroductionAgrobacterium-mediated genetic transformation has been widely used for the identification of functional genes and regulatory and developmental mechanisms in plants. However, there are still some problems of low genetic transformation efficiency and high genotype dependence in cruciferous crops.MethodsIn this study, broccoli, a worldwide Brassica crop, was used to investigate the effects of genotype, explant type, concentration of hygromycin B used during seedling selection, overexpression vector type, RNAi and CRISPR/cas9 on the genetic transformation efficiency. At the same time, two vectors, PHG-031350 and PHG-CRa, were used for subcellular localization of the glucoraphanin synthesis-related gene FMOGS-OX5 and clubroot resistance gene by a PEG-Ca2+-mediated transient transformation system for broccoli protoplasts. Finally, the Agrobacterium-mediated genetic transformation system of broccoli was optimized and improved.Results and DiscussionThis study showed that hypocotyl explants are more suitable for Agrobacterium-mediated transgene and CRISPR/Cas9 gene editing of broccoli. In contrast to previous studies, we found that 5 mg/L hygromycin B was more advantageous for the selection of resistant broccoli sprouts, and genotype 19B42 reached the highest transformation rate of 26.96%, which is higher than that in Brassica oleracea crops. In addition, the inbred line 19B42 successfully achieved high genetic transformation of overexpression, RNAi and CRISPR/Cas9 vectors; thus, it is powerful recipient material for the genetic transformation of broccoli. Subcellular localization proved that the glucoraphanin metabolism-related gene Bol031350 and clubroot resistance gene CRa were both expressed in the cytoplasm and nucleus, which provided a scientific basis for studying the regulation of glucosinolate metabolism and clubroot resistance in cruciferous crops. Therefore, these findings will provide new insight into the improvement of the genetic transformation and molecular breeding of Brassica oleracea crops.

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Section: Introductionmentioning

confidence: 99%

A highly efficient genetic transformation system for broccoli and subcellular localization

et al. 2023

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“…In general, a large amount of soluble protein in plant biomass can provide better production of recombinant proteins, thereby enhancing the possibility of an efficient plant-based bioreactor system [10]. Chinese cabbage transformation protocols are well established for the overexpression of transgenes, which can impart resistance to plant pathogenesis [11][12][13] and tolerance to harsh environments, such as drought and salinity [14][15][16], and it can also improve physiological traits, such as tolerance to calcium deficiency [17]. There are several advantages of using Chinese cabbage for the production of therapeutic proteins.…”

Section: Introductionmentioning

confidence: 99%

Expression of Colorectal Cancer Antigenic Protein Fused to IgM Fc in Chinese Cabbage (Brassica rapa)

Lee

Lim

et al. 2020

Plants

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The epithelial cell adhesion molecule (EpCAM) is a tumor-associated antigen and a potential target for tumor vaccine. The EpCAM is a cell-surface glycoprotein highly expressed in colorectal carcinomas. The objective of the present study is to develop an edible vaccine system through Agrobacterium-mediated transformation in Chinese cabbage (Brassica rapa). For the transformation, two plant expression vectors containing genes encoding for the EpCAM recombinant protein along with the fragment crystallizable (Fc) region of immunoglobulin M (IgM) and Joining (J)-chain tagged with the KDEL endoplasmic reticulum retention motif (J-chain K) were constructed. The vectors were successfully transformed and expressed in the Chinese cabbage individually using Agrobacterium. The transgenic Chinese cabbages were screened using genomic polymerase chain reaction (PCR) in T0 transgenic plant lines generated from both transformants. Similarly, the immunoblot analysis revealed the expression of recombinant proteins in the transformants. Further, the T1 transgenic plants were generated by selfing the transgenic plants (T0) carrying EpCAM–IgM Fc and J-chain K proteins, respectively. Subsequently, the T1 plants generated from EpCAM–IgM Fc and J-chain K transformants were crossed to generate F1 plants carrying both transgenes. The presence of both transgenes was validated using PCR in the F1 plants. In addition, the expression of Chinese cabbage-derived EpCAM–IgM Fc × J-chain K was evaluated using immunoblot and ELISA analyses in the F1 plants. The outcomes of the present study can be utilized for the development of a potential anti-cancer vaccine candidate using Chinese cabbage.

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“…Aphids nymphs (without wings) were collected from infected wild type B. juncea plant grown in field condition were used in the experiment. Aphid movements were recorded on transgenic leaves emitting Ebf using detached leaf 11,12 for aphid bioassay. A circular disc from the leaves of WT plants (2 cm in diameter) was removed, and replaced by a similar size disc excised from the leaves of transgenic plants.…”

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(E)-β-farnesene gene reduces Lipaphis erysimi colonization in transgenic Brassica juncea lines

Verma

Sinha²,

Jajoo

2015

Plant Signaling & Behavior

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Aphids are the major concern that significantly reduces the yield of crops. (E)-b-farnesene (Ebf) is the principal component of the alarm pheromone of many aphids. The results of current research support the direct defense response of (E)-b-farnesene (Ebf) against aphid Lipaphis erysimi (L.) Kaltenbach in Brassica juncea. Ebf gene was isolated from Mentha arvensis and transformed into B. juncea, showed direct repellent against aphid colonization. The seasonal mean population (SMP) recorded under field condition showed significantly higher aphid colonization in wild type in comparison to most of the transgenic lines, and shows positive correlation with the repellency of transgenic plant expressing (E)-b-farnesene. The current research investigation provides direct evidence for aphid control in B. juncea using Ebf, a non-toxic mode of action.

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High resistance of transgenic cabbage plants with a synthetic cry1Ia8 gene from Bacillus thuringiensis against two lepidopteran species under field conditions

Abstract: Compared with the control, the homozygous transgenic cabbage line showed great potential for protecting cabbage from attack by P. xylostella and P. rapae in the field.

Cited by 7 publications

References 19 publications

A highly efficient genetic transformation system for broccoli and subcellular localization

A highly efficient genetic transformation system for broccoli and subcellular localization

Expression of Colorectal Cancer Antigenic Protein Fused to IgM Fc in Chinese Cabbage (Brassica rapa)

(E)-β-farnesene gene reduces Lipaphis erysimi colonization in transgenic Brassica juncea lines

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