[Objective] To cultivate blanket rapeseed seedlings suitable for automatic mechanical transplanting, the effects of seed soaking with uniconazole and substrate formula on seed germination and seedling quality of rapeseed were investigated using HY4 as the material. [Method] The rapeseed seeds were soaked in different concentrations(0, 5, 10, 20, 30, 40 and 50 mg/L) of uniconazole for 0, 1, 2, 4 and8 h, respectively. Subsequently, they were dried naturally. After the preliminary screening carried out in petri dishes and in a laboratory, the optimum soaking concentration and time of uniconazole were obtained. On the other hand, the rapeseed seeds were soaked in different concentrations(0, 5, 10, 15 and 20 mg/L) of uniconazole for 0, 1 and 2 h, respectively for outdoor pot experiment. At the same time, the screening of optimum substrate formula for cultivating blanket rapeseed seedlings was carried out. The field test was carried out to verify the screened optimum substrate formula. [Result] The results of petri dish experiment showed that seed soaking with uniconazole could delay seed germination, reduce seed germination rate, inhibit hypocotyl elongation and promote root growth. When the soaking concentration exceeded 30 mg/L and soaking time exceeded 2 h, the effect of seed soaking with uniconazole on seed germination was greater, and the seed soaking inhibited root growth. The results of outdoor pot experiment showed that the soaking concentrations and times all significantly reduced seedling height, slowed shoot growth and promoted root growth, which all contributed to the cultivation of blanket rapeseed seedlings suitable for automatic mechanical transplanting. In the substrate formula, the lower the proportion of soil was, the better the seedling growth was. In the substrate with lower proportion of soil, the seed germination rate, plant height,root vigor and dry matter accumulation were all increased(P〈0.05), which was verified by the field test. When the soaking concentration was less than 10
This study was to investigate the efficiency and specificity of RNAi silencing on the expression of endogenous fad2 gene in transgenic line W-4. [Method] The relative expression of fad2 gene in seeds at different developmental stages of 7th, 14th, 21st and 28th day after flowering (DAF) as wel as the root, stem, leaf at winter seedling stages of both the transgenic line W-4 and non-transgenic control Westar by real-time fluorescence quantitative PCR. [Results] The results showed the relative expression of fad2 gene was gradual y increasing with the days after flowering in the seeds of the control Westar, while it was found decreasing significantly since the 21st DAF in the seeds of the line W-4. The decline was up to 60% in comparison with the control Westar. However, no significant difference in the relative expression of fad2 gene in other organs like root, stem and leaf was observed between transgenic line W-4 and non-transgenic control Westar. Fatty acid composition analysis showed the oleic acid desaturation parameter(ODP) in seeds of the line W-4 was 0.07 in average, decreased by nearly 75% than control Westar which was 0.24 in average, while no significant difference in the seedling root, stem and leaf was measured between transgenic rapeseed and control. [Conclusion] The results above validated that RNA interference in transgenic rapeseed W-4 is at a seed-specific manner, not interfering with fad2 gene expression in organs such as the root, stem and leaf. The study also found that the period of fad2 gene expres-sion decline was wel coincided with the expression of napin gene, both appeared at the 21st DAF, indicating that the expression of dsRNA of fad2 gene is precisely control ed by the napin promoter.
A predominantly expressed gene, pyruvate kinease (PK) gene, control ing oil accumulation, had been identified in previous study. To construct a PK RNAi vector, a 498-bp target PK gene sequence was amplified and transferred into the pEASY-T1vector. Subsequently, the target DNA fragments were cut off by enzymes Not I and Xho I and directional y inserted into plant RNAi platform vector pHurricane, a newly developed RNAi vector in our lab, to form the PK inverse repeats. The PK inverted repeats fragment was then cloned into a modified vector pCAMBIA1390, driven by a rapeseed seed-specific promoter napin. Restriction enzyme digestion verified the successful construction of RNA interference vector. The PK RNAi con-struction wil lay a foundation for study in the function of PK in oil accumulation and metabolism in rapeseeds.