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Acta Agronomica Sinica ›› 2018, Vol. 44 ›› Issue (12): 1829-1836.doi: 10.3724/SP.J.1006.2018.01829

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Spatio-temporal Expression of Bt Protein and Stem Borer Resistance of Transgenic Early Japonica Rice with cry1C* or cry2A* Gene

Rong-Tian LI1,Xin-Yu WANG1,Chong-Bing TIAN1,Qing ZHOU1,Chang-Hua LIU1,2,*()   

  1. 1 Key Laboratory of Molecular Biology, College of Heilongjiang Province / Heilongjiang University, Harbin 150080, Heilongjiang, China
    2 College of Agricultural Resources and Environment, Heilongjiang University, Harbin 150080, Heilongjiang, China
  • Received:2018-01-23 Accepted:2018-08-20 Online:2018-12-12 Published:2018-09-29
  • Contact: Chang-Hua LIU E-mail:liuchanghua70@163.com
  • Supported by:
    This study was supported by the National Major Project for Developing New GM Crops(2016ZX08001001-001-007);the National Major Project for Developing New GM Crops(2014AA10A600);the Open Project of Key Laboratory of Molecular Biology, College of Heilongjiang Province

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Abstract:

The early japonica rice Kongyu 131 (Oryza sativa) was transformed with cry1C* or cry2A* gene driven by ubi promoter using Agrobacterium tumefaciens mediated method to create the transgenic early japonica rice Kongyu 131 (cry1C*) and Kongyu 131 (cry2A*). The experiments were conducted by transplanting the different lines of Kongyu 131 (cry1C*) and Kongyu 131 (cry2A*) from independent transformation events in the field to detect Bt protein content (BPC) of organs of the transgenic rice at different growth stages with enzyme-linked immune sorbent assay (ELISA) and resistance to rice striped stem borer as evidenced by insect feeding bioassays. The BPC of cry1C* gene was always lower than that of cry2A* in early japonica rice. There was a difference in BPC at different growth stages of transgenic rice, displaying tillering stage < heading stage < filling stage in leaf blade or sheath-stem and heading stage > filling stage > maturity in young panicle or brown rice. BPC of organs of transgenic rice was different at every stage, the order of BPC from high to low was leaf blade and sheath-stem at tillering stage, leaf blade, young panicle and sheath-stem at heading stage, leaf blade, sheath-stem, young panicle and brown rice at filling stage and maturity. There were some obvious differences in BPC of leaf blade, sheath-stem and young panicle at heading stage or brown rice at maturity and in borer-resistant property among transgenic lines with cry1C* or cry2A*. Correlations between BPC of each organ at heading stage and borer-resistant property or BPC of brown rice were not found in transgenic rice lines with cry1C* or cry2A*. The BPC increased in vegetative organs while descended in reproductive organs in the process of growth and development, and BPC of vegetative organs was usually higher than BPC of reproductive organs in both of Kongyu 131 (cry1C*) and Kongyu 131 (cry2A*). All transgenic early japonica rice showed effective resistance to stem borer in the experiment, though there was significant difference in BPC either between cry1C* and cry2A* genes or among transgenic rice lines.

Key words: cry1C* gene, cry2A* gene, transgenic early japonica rice, Bt protein content, spatio-temporal expression, stem borer resistance

Fig. 1

T-DNA region of Ti plasmid pBar13-cry1C*(A) and pBar13-cry2A*(B) RB: right border; ubi: ubi promoter; cry1C*: cry1C * gene; cry2A*: cry2A * gene; 35S: CaMV35S promoter; bar: phosphinothricin acetyltransferase gene; LB: left border."

Fig. 2

Bt protein content of HD1 and HD2 of transgenic early japonica rice A: leaf blade; B: sheath-culm; C: young panicle or brown rice; BPC: Bt protein content. *, **: significant at P < 0.05 and P < 0.01, respectively."

Fig. 3

Bt protein content of the transgenic early japonica rice among the different growth stages A: leaf blade; B: sheath-culm; C: young panicle or brown rice; BPC: Bt protein content. *, **: significant at P < 0.05 and P < 0.01, respectively."

Fig. 4

Bt protein content of the transgenic early japonica rice at different growth stages A: tillering stage; B: heading stage; C: filling stage and maturity; BPC: Bt protein content. **: significant at P < 0.01."

Table 1

Bt protein content and resistance to stem borer in different lines of transgenic early japonica rice"

品系
Line		 叶片Bt蛋白量
BPC of leaf blade (μg g-1)		 茎鞘Bt蛋白量
BPC of sheath-culm
(μg g-1)		 幼穗Bt蛋白量
BPC of young panicle (μg g-1)		 糙米Bt蛋白量
BPC of brown rice (μg g-1)		 校正幼虫死亡率
Adjusted larval mortality rate (%)
HD1-1 5.33±0.04** 0.50±0.03** 0.52±0.02** 0.09±0.01** 96.46±1.42**
HD1-2 2.74±0.03** 0.49±0.02** 0.51±0.03** 0.15±0.02** 91.34±1.02**
HD1-3 1.16±0.02** 0.44±0.03** 0.43±0.03** 0.06±0.02** 92.08±2.70**
HD1-4 1.16±0.02** 0.31±0.03** 0.36±0.02** 0.10±0.02** 96.20±1.06**
HD2-1 21.81±0.02** 2.87±0.04** 3.95±0.02** 0.33±0.03** 94.21±1.05**
HD2-2 29.65±0.02** 5.69±0.02** 4.73±0.02** 0.24±0.02** 93.73±3.55**
HD2-3 21.63±0.04** 3.63±0.02** 4.02±0.05** 0.55±0.03** 91.56±4.10**
HD2-4 49.63±0.06** 4.64±0.02** 5.43±0.04** 0.13±0.03** 98.06±0.88**

Table 2

Relationship of Bt protein content of the organs at heading stage and brown rice with insect-resistance among the lines of transgenic early japonica rice"

转基因早粳稻
Transgenic early japonica rice		 抽穗期性状
Characteristic at heading stage		 成熟期糙米Bt蛋白量
BPC of brown rice at maturity		 校正幼虫死亡率
Adjusted larval mortality rate
HD1 叶片Bt蛋白量 BPC of leaf blade 0.233 0.375
茎鞘Bt蛋白量 BPC of sheath-culm 0.254 -0.389
幼穗Bt蛋白量 BPC of young panicle 0.381 -0.267
HD2 叶片Bt蛋白量 BPC of leaf blade -0.796 0.910*
茎鞘Bt蛋白量 BPC of sheath-culm -0.533 0.250
幼穗Bt蛋白量 BPC of young panicle -0.827 0.838
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