本文選題：能源植物 + 甜高粱�。� 參考：《江蘇大學》2017年碩士論文

【摘要】：甜高粱是一種C4植物,由于它生長速度快,產(chǎn)量高,含糖量高,且具有抗逆性強的特點,適宜在鹽堿灘涂及干旱等邊際土壤大規(guī)模種植。研究甜高粱的耐鹽性,篩選耐鹽品種并對其進行遺傳改良,對于緩解我國糧食壓力及資源化利用邊際土壤,具有十分重要的意義。本研究首先從不同濃度的鹽脅迫(NaCl)對四個甜高粱品種(系)大力王(DLW)、牛魔王(NMW)、海牛(HN)和帕卡(PK)種子萌發(fā)和幼苗生長等方面的影響,對甜高粱發(fā)芽期耐鹽性進行初步比較與評價。研究表明NMW相對其他品種(系)的耐鹽性最強,其次是DLW,而PK和HN易受鹽脅迫的影響。研究表明當鹽脅迫濃度較低(80 m M)時,保護酶活性升高及滲透壓調(diào)節(jié)物質(zhì)的增加有利于增強甜高粱的耐鹽性,而當鹽脅迫濃度過高(240 m M)時,植物的保護酶系統(tǒng)產(chǎn)生不可逆破壞,此時滲透壓調(diào)節(jié)物質(zhì)對保護植物起到重要的作用。以甜高粱成熟種子為外植體建立甜高粱組織培養(yǎng)體系的研究發(fā)現(xiàn):誘導甜高粱愈傷組織時2,4-D最佳濃度為4 mg/L,而IAA和6-BA的添加會降低愈傷組織的誘導率及胚性愈傷組織產(chǎn)生。四個甜高粱品種(系)中DLW的愈傷組織誘導效果最佳,其次為NMW,而HN和PK的愈傷組織誘導效果較差。以DLW為外植體進行愈傷組織的再分化及生根實驗得出:最佳的愈傷組織分化培養(yǎng)基為MS+4 mg/L 6-BA+0.5 mg/L IBA,不定根誘導培養(yǎng)基為MS+3 mg/L IAA。本實驗成功建立了DLW的高效組織培養(yǎng)體系,為DLW遺傳改良的研究奠定了基礎(chǔ)。以甜高粱品種DLW的愈傷組織為外植體,對甜高粱遺傳轉(zhuǎn)化過程中乙酰丁香酮濃度、潮霉素篩選濃度、農(nóng)桿菌菌液濃度、真空浸染時間、共培養(yǎng)時間等條件進行優(yōu)化,通過農(nóng)桿菌介導的植物遺傳轉(zhuǎn)化法成功將新疆鹽穗木的耐鹽堿基因HcNHX1轉(zhuǎn)入甜高粱品種DLW中。DLW遺傳轉(zhuǎn)化的最佳條件為:100μM乙酰丁香酮,菌液濃度OD600≈0.6,抽真空5 min,共培養(yǎng)時間3 d,潮霉素篩選濃度為20 mg/L。通過對轉(zhuǎn)基因抗性植株進行分子水平檢測,表明本研究共獲得5株T0代含有功能基因HcNHX1的轉(zhuǎn)基因植株。對轉(zhuǎn)基因植株與野生型植株的根尖細胞活力、根尖活性氧及植株的Na+、K+含量進一步比較的研究表明,與野生型植株相比,轉(zhuǎn)基因植株具有較高的根尖細胞活力、較少的根尖活性氧含量,以及Na+含量,說明獲得的轉(zhuǎn)基因植株對鹽脅迫具有較好的抗性。
[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，

本文編號：1801694