To more effectively control two major cotton insects (cotton bollworm and Spodoptera litura) and improve the efficacy of the pest resistance management, novel transgenic plants expressing Bacillus thuringiensis Cry9C gene were generated, and gene stacking strategy was incorporated. Initially, a binary plasmid vector harboring Cry9C gene was introduced into an elite cotton cultivar Simian-3 by Agrobacterium-mediated transformation. Integration and expression of the Cry9C genes in three transgenic lines were confirmed by PCR and RT-PCR. Among these transgenic lines, T0 generation of line 16 (L-16) with normal phenotypes were selected by ELISA assays for its highest expression level of Cry9C. In T1 population of L-16, the expression level of Cry9C ranged from 29 to 45 lg/g fresh leaf. The following insect bioassays demonstrated that transgenic S3-35S::Cry9C cotton plants exhibited moderate toxicity to Heliothis armigera but strong toxicity to S. litura compared with the transgenic plants expressing Cry 1Ac gene. For incorporation of gene staking strategy, Cry9C gene and Cry 2A or Cry 1Ac were pyramided, respectively by sexual crossing. The expression of Cry9C protein in all F1 progenies had a similar level as the parent plants indicating the high heritability of Bt genes in transgenic progenies. Progenies from both Cry9C × Cry 2A and Cry9C × Cry 1Ac exhibited higher resistance to S. litura compared with their parents. Together our data demonstrated that our newly generated transgenic plants represent a reservoir of novel insect-resistant materials in cotton breeding, and the successful incorporation of gene pyramiding technology can provide a new solution of developing multiple resistance management strategies.
In this study, the effects of salicylic acid to antioxidative activity and photosynthetic characteristics in waterlogging stress of two peony cultivars (‘Fengdanbai’ and ‘Mingxing’) were investigated. 4-year-old peony grown in different levels of waterlogging stress and then different concentration prepared SA (0.0, 0.1, 0.5 and 1.0 mmol L⁻¹) sprayed on fresh leaves of peony. The antioxidative enzymes activities include superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT), chlorophyll content, relative conductivity and MDA content were measured in leaves about different waterlogging treatment, the photosynthetic characteristics were also measured using photosynthetic measurement system. The results showed that waterlogging stress decreased the chlorophyll content in all peony cultivars leaves, but with SA treatment can inhibit the decrease of chlorophyll content. Relative conductivity increased as the extension of waterlogging time in two cultivars. SA treatment could effectively inhibit the increase of relative conductivity, and 0.5 mmol L⁻¹ of SA was the most suitable concentration. SOD, POD, CAT activity increased first and then decreased in different waterlogging condition, SA significantly increased the activity of various enzymes. MDA content was increase as the expansion of waterlogging time in two cultivars. SA inhibits the increase of MDA content. Of all concentration of SA, 0.5 mmol L⁻¹ was the best concentration to inhibit the waterlogging stress. For the photosynthetic characteristics, the net assimilation rate (Pn), stomatal conductance (Gs), transpiration rate (Tr) and intercellular CO₂ (Ci) were decreased under different waterlogging condition. SA treatment can increase Pn, Gs, Tr and Ci of peony.