能源植物甜高粱耐盐品种的筛

发布时间：2018-04-25 14:24

  本文选题：能源植物 + 甜高粱　； 参考：《江苏大学》2017年硕士论文

【摘要】：甜高粱是一种C4植物,由于它生长速度快,产量高,含糖量高,且具有抗逆性强的特点,适宜在盐碱滩涂及干旱等边际土壤大规模种植。研究甜高粱的耐盐性,筛选耐盐品种并对其进行遗传改良,对于缓解我国粮食压力及资源化利用边际土壤,具有十分重要的意义。本研究首先从不同浓度的盐胁迫(NaCl)对四个甜高粱品种(系)大力王(DLW)、牛魔王(NMW)、海牛(HN)和帕卡(PK)种子萌发和幼苗生长等方面的影响,对甜高粱发芽期耐盐性进行初步比较与评价。研究表明NMW相对其他品种(系)的耐盐性最强,其次是DLW,而PK和HN易受盐胁迫的影响。研究表明当盐胁迫浓度较低(80 m M)时,保护酶活性升高及渗透压调节物质的增加有利于增强甜高粱的耐盐性,而当盐胁迫浓度过高(240 m M)时,植物的保护酶系统产生不可逆破坏,此时渗透压调节物质对保护植物起到重要的作用。以甜高粱成熟种子为外植体建立甜高粱组织培养体系的研究发现:诱导甜高粱愈伤组织时2,4-D最佳浓度为4 mg/L,而IAA和6-BA的添加会降低愈伤组织的诱导率及胚性愈伤组织产生。四个甜高粱品种(系)中DLW的愈伤组织诱导效果最佳,其次为NMW,而HN和PK的愈伤组织诱导效果较差。以DLW为外植体进行愈伤组织的再分化及生根实验得出:最佳的愈伤组织分化培养基为MS+4 mg/L 6-BA+0.5 mg/L IBA,不定根诱导培养基为MS+3 mg/L IAA。本实验成功建立了DLW的高效组织培养体系,为DLW遗传改良的研究奠定了基础。以甜高粱品种DLW的愈伤组织为外植体,对甜高粱遗传转化过程中乙酰丁香酮浓度、潮霉素筛选浓度、农杆菌菌液浓度、真空浸染时间、共培养时间等条件进行优化,通过农杆菌介导的植物遗传转化法成功将新疆盐穗木的耐盐碱基因HcNHX1转入甜高粱品种DLW中。DLW遗传转化的最佳条件为:100μM乙酰丁香酮,菌液浓度OD600≈0.6,抽真空5 min,共培养时间3 d,潮霉素筛选浓度为20 mg/L。通过对转基因抗性植株进行分子水平检测,表明本研究共获得5株T0代含有功能基因HcNHX1的转基因植株。对转基因植株与野生型植株的根尖细胞活力、根尖活性氧及植株的Na+、K+含量进一步比较的研究表明,与野生型植株相比,转基因植株具有较高的根尖细胞活力、较少的根尖活性氧含量,以及Na+含量,说明获得的转基因植株对盐胁迫具有较好的抗性。
[Abstract]:Sweet sorghum is a kind of C4 plant. Because of its high growth rate, high yield, high sugar content and strong resistance to stress, sweet sorghum is suitable for large-scale planting in marginal soils such as saline beach and drought. It is very important to study the salt tolerance of sweet sorghum, to screen salt-tolerant varieties and to make genetic improvement on them for relieving grain pressure and utilizing marginal soil in China. In this study, the effects of different concentrations of salt stress on seed germination and seedling growth of four sweet sorghum varieties (lines), DLWN, NMWF, HNN and PKK, were compared and evaluated. The results showed that NMW had the strongest salt tolerance than other varieties (lines), followed by DLW, while competition and HN were susceptible to salt stress. The results showed that the increase of protective enzyme activity and osmotic pressure regulator was beneficial to enhance the salt tolerance of sweet sorghum when the concentration of salt stress was 80 mm 路m ~ (-1), but when the concentration of salt stress was too high, the salt tolerance of sweet sorghum was increased, but when the concentration of salt stress was too high, the salt tolerance of sweet sorghum was increased. The protective enzyme system of plants is irreversibly destroyed, and osmotic pressure regulators play an important role in protecting plants. The study on the establishment of sweet sorghum tissue culture system by using mature seeds of sweet sorghum as explant showed that the optimum concentration of 24-D was 4 mg / L when inducing sweet sorghum callus, while the addition of IAA and 6-BA could reduce the induction rate of callus and the formation of embryogenic callus. Among the four sweet sorghum varieties (lines), DLW had the best callus induction effect, followed by NMWs, but HN and competing callus induction was poor. The experiment of callus redifferentiation and rooting with DLW as explant showed that the best medium for callus differentiation was MS 4 mg/L 6-BA 0.5 mg/L IBA, and the adventitious root induction medium was MS 3 mg/L IAA. In this experiment, the efficient tissue culture system of DLW was successfully established, which laid a foundation for the study of genetic improvement of DLW. The callus of sweet sorghum variety DLW was used as explant to optimize the concentration of acetyl eugenone, hygromycin, Agrobacterium tumefaciens, vacuum soaking time and co-culture time during genetic transformation of sweet sorghum. By means of Agrobacterium tumefaciens mediated plant genetic transformation, the saline-tolerant gene HcNHX1 of Xinjiang salt panicle was successfully transferred into sweet sorghum variety DLW. The optimum condition for genetic transformation was: 1. 100 渭 M acetyl eugenone. The concentration of OD600 鈮，

本文编号：1801694