Identification and Analysis of Important Phenotypes of E1C608 with Glyphosate Resistance and Lepidopteran Resistance in Rice

doi:10.13523/j.cb.20191104

China Biotechnology

2019, Vol. 39

Issue (11): 31-38 DOI: 10.13523/j.cb.20191104

Identification and Analysis of Important Phenotypes of E1C608 with Glyphosate Resistance and Lepidopteran Resistance in Rice

ZENG Qiang^1,²,MENG Qiu-cheng¹,DENG Li-hua¹,LI Jin-jiang¹,YU Jiang-hui¹,WENG Lu-shui¹,XIAO Guo-ying^1,³(

)

1 Key Laboratory of Agro-ecological Processes in Subtropical Region, Institute of Subtropical Agriculture,Chinese Academy of Sciences, Changsha 410125, China
2 University of Chinese Academy of Sciences, Beijing 100049, China

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Abstract

Weeds and insect pests are two important biotic stresses to result in yield loss in rice, and breeding bioengineered rice with herbicide resistance and insect resistance is one of the effective ways to solve these two problems. And for that, the glyphosate resistance and Lepidopteran resistance as well as molecular and some agronomic characteristics of transgenic line E1C608 in rice were identified. Results of molecular identification showed that three independent transformation events were obtained, in which one single copy insertion event was named as E1C608-3. The transformant E1C608-3 of T₂ generation was used to detect exogenous protein contents in root, stem and leaf at tillering stage by ELISA. The protein contents of EPSPS and CRY1C in different organs of E1C608-3 were significantly different both in descending order of leaf>stem>oot (P<0.01), ranged from120.16μg/g to 1 223.28μg/g and from 1.23μg/g to 8.72μg/g, respectively. The glyphosate tolerable concentration at seedling stage of E1C608-3 in T₃ generation reached at least 16g/L, which was sixteen times higher than that of the transformation receptor R608. The larva mortality of rice leaf roller fed on leaves of E1C608-3 in T₃ generation for five days was 95.56%, that showed the E1C608-3 with an excellent resistance to Lepidopteran insects. The data suggested that some agronomic traits were significantly different between E1C608-3 in T₄ generation and R608 (P<0.01), but all of the variations were within the variation range of natural germplasm resource in rice. Overall, a novel germplasm with glyphosate resistance and Lepidopteran resistance was verified to be developed in rice.

Key words： Rice Bioengineering Glyphosate resistance Lepidopteran resistance Agronomic traits

Received: 28 March 2019 Published: 17 December 2019

ZTFLH:

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Corresponding Authors: Guo-ying XIAO E-mail: xiaoguoying@isa.ac.cn

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	Qiang ZENG
	Qiu-cheng MENG
	Li-hua DENG
	Jin-jiang LI
	Jiang-hui YU
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	Guo-ying XIAO

Cite this article:

ZENG Qiang,MENG Qiu-cheng,DENG Li-hua,LI Jin-jiang,YU Jiang-hui,WENG Lu-shui,XIAO Guo-ying. Identification and Analysis of Important Phenotypes of E1C608 with Glyphosate Resistance and Lepidopteran Resistance in Rice. China Biotechnology, 2019, 39(11): 31-38.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20191104 OR https://manu60.magtech.com.cn/biotech/Y2019/V39/I11/31

Fig. 1 The T-DNA region of plant expression vector pC3300-Epsps-35SCry1Ca

Table 1 The primers for PCR and Southern blot

Fig. 2 Molecular identification of T₁ transgenic plants (a) PCR detection (b) Southern blotting analysis M: DNA molecular weight marker; +: Plasmid pC3300-Epsps-35SCry1Ca; EcoRI and HindIII: Restriction enzymes; R608: Transformation receptor; 1, 2 and 3: Transformant E1C608-1, E1C608-2, E1C608-3, respectively

Fig. 3 Quantification of EPSPS and CRY1C proteins (a) Comparison of EPSPS protein contents in root, stem and leaf of E1C608-3 (b) Comparison of CRY1C protein contents in root, stem and leaf of E1C608-3 Different capital or small letters mean significant difference at 0.01 or 0.05 level, respectively; and the error bar represents standard deviation

Fig. 4 Spraying glyphosate at different concentrations on T₃ generation transgenic plants E1C608-3 is a transgenic line; R608 is of wild type; 0, 0.5g/L, 1g/L, 2g/L, 3g/L, 8g/L, 10g/L, 12g/L, 14g/L and 16g/L indicates different concentrations of glyphosate solution

Fig. 5 Evaluation of resistance to rice leaf roller (a)Leaves gnawed by rice leaf rollers for five days (b) Mortality of rice leaf rollers in five days E1C608 is of single-copy insertion events in T₃ generation; R608 is of wild type. Different capital or small letters mean significant difference at 0.01 or 0.05 level, respectively, and the error bar represents standard deviation

Table 2 Agronomic traits and their classifications of transgenic rice E1C608 in T₄ generation


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