转JERF36基因银中杨的抗旱性评价及生理机理研究

发布时间：2018-04-16 10:10

本文选题：干旱胁迫 + 转基因银中杨　；参考：《中国林业科学研究院》2016年硕士论文

【摘要】：杨树(Populus)是世界上分布最广、栽培面积最大的速生用材林树种之一,也是我国北方地区最重要的造林树种。长期以来,杨树遗传改良以追求速生性为主,而忽视其抗逆性,致使其在我国广大的干旱、半干旱地区难以发挥应有作用。利用基因工程开展杨树抗旱品种选育和改良,对干旱、半干旱荒地利用将产生积极作用。本文以转JERF36基因银中杨(P.alba×P.berolinensis)(ABJ01)和非转基因银中杨(9#)为试验材料,通过测定土壤控水干旱胁迫条件下植株生长、叶片解剖结构、光合参数、生理生化指标,和水培PEG模拟干旱胁迫条件下根尖离子流等指标,开展了转基因银中杨的抗旱性评价研究,初步揭示转JERF36基因银中杨抗旱性生理机制,为进一步深入研究转基因植物的抗旱性机理奠定基础,并为转基因杨树的推广应用提供科学依据。主要研究结果如下:1.转JERF36银中杨在干旱胁迫下生长优势明显,叶片栅栏组织发达,其抗旱能力得到提高。干旱胁迫处理的3个时间点上(10 d、20 d、30 d),ABJ01的苗高、地径均大于9#,中度干旱胁迫处理20 d时,ABJ01的地径显著高于9#,重度干旱胁迫下处理20 d和30 d时,ABJ01的苗高显著高于9#;中度和重度干旱处理30 d时,ABJ01的单叶面积和总干重显著大于9#。中度干旱胁迫处理20 d时,ABJ01的叶片厚度显著高于9#;重度干旱胁迫下,在3个处理时间点上ABJ01的叶片上、下表皮厚度均显著高于9#;中度干旱胁迫处理10 d和30 d时及重度干旱胁迫处理20 d时,ABJ01栅栏组织厚度显著高于9#;中度和重度干旱胁迫处理30 d时,ABJ01的海绵组织厚度显著低于9#。2.干旱胁迫下,转JERF36银中杨较非转基因银中杨具有更强的光合能力、气体交换能力和保水能力,且随着干旱胁迫程度的增加,差异越明显。中度干旱胁迫处理20 d和重度干旱胁迫下3个处理时间点上,ABJ01的净光合速率(Pn)和气孔导度(Gs)均显著高于9#;中度和重度干旱胁迫处理20 d和30 d时,ABJ01的蒸腾速率(Tr)低于9#,ABJ01的叶绿素a、叶绿素b和总叶绿素含量较9#高,重度干旱胁迫条件下达到显著水平。中度和重度干旱胁迫处理30 d时,ABJ01的Fv/Fm值较9#高,中度胁迫条件下达显著水平,表明ABJ01的光系统受损情况较9#小。3.与非转基因银中杨相比,转JERF36银中杨在干旱胁迫下的渗透调节能力和活性氧清除能力更强。中度和重度干旱胁迫处理20 d和30 d时,ABJ01的脯氨酸(Pro)含量均比9#高,中度干旱胁迫下达到显著水平。中度和重度干旱胁迫条件下,处理20 d和30 d时,ABJ01叶片的超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性及过氧化氢酶(CAT)活性均高于9#,差异均达显著水平。4.PEG胁迫条件下,转基因株系根系具有更强的K~+、Ca~(2+)吸收能力。PEG胁迫下ABJ01的K~+外排流速低于9#,10%PEG胁迫下达到显著水平;20%PEG胁迫下,ABJ01的根尖Ca~(2+)内流能力显著高于9#。20%PEG胁迫下ABJ01的根中K元素显著高于9#,10%和20%PEG胁迫下ABJ01的根系能够积累更多的Ca元素。总体上,转基因银中杨在干旱胁迫下生长优势明显。其抗旱生理机制主要体现在通过增加栅栏组织厚度、降低海绵组织厚度、提高渗透调节和活性氧清除能力,减少蒸腾等响应来提高其干旱胁迫适应能力,同时转基因银中杨在干旱胁迫下Pn以及气体交换能力得到有效提高,维持快速生长的能力。另外,外源基因JERF36的导入能够增加转基因银中杨根尖胞质Ca~(2+)内流,减少K~+的损失,维持根尖较高的Ca~(2+)、K~+水平,以增强转基因银中杨的渗透调节能力和吸水能力,来增强转基因银中杨的对干旱胁迫的耐受能力,从而提高转基因银中杨的抗旱能力。
[Abstract]:Poplar (Populus) is the world's most widely distributed, the largest cultivated area of fast-growing timber species is one of the most important tree species in northern area of China. For a long time, poplar genetic improvement in pursuit of rapid growth, while ignoring its resistance, resulting in China's vast arid and semi-arid regions, it is difficult to play role. The use of genetic engineering to carry out breeding and improvement of poplar varieties to drought, drought, will have a positive effect in semi arid wasteland. This paper use the JERF36 transgenic poplar (P.alba * P.berolinensis) (ABJ01) and non transgenic poplar (9#) as the experimental material, the growth through the determination of soil water control drought stress plant, leaf anatomical structure, photosynthetic characteristics, physiological and biochemical indexes, and hydroponic PEG simulation under drought stress conditions of apical ion flow and other indicators, carried out research on the evaluation of drought resistance of transgenic poplar, revealed to Yang drought resistance physiological mechanism of JERF36 gene in silver, and lay the foundation for further research on the mechanism of drought resistance of transgenic plants, and for the application of transgenic poplar and provide a scientific basis. The main results are as follows: 1. JERF36 transgenic poplar under drought stress growth advantage, palisade tissue developed, the drought resistance ability is improved. 3 at the time of drought stress on (10 d, 20 D, 30 d ABJ01), the seedling height, ground diameter was greater than 9#, moderate drought stress of 20 D, ABJ01 diameter was significantly higher than that of 9#, under severe drought stress treatment for 20 D and 30 d ABJ01, the seedling height significantly 9# is higher than that of moderate and severe drought treatment; 30 d, ABJ01 of the total dry weight and leaf area were significantly higher than that of 9#. under moderate drought treatment for 20 D, the thickness of leaf ABJ01 was significantly higher than that of 9#; under severe drought stress, the leaf ABJ01 in the 3 time points of upper and lower epidermis thickness The degree was significantly higher than that of 9#; moderate drought stress of 10 D and 30 d and severe drought stress of 20 d ABJ01, the thickness of palisade tissue was significantly higher than that of 9#; moderate and severe drought stress of 30 d, the thickness of spongy tissue ABJ01 was significantly lower than that of 9#.2. under drought stress, transgenic JERF36 poplar Jiaofei transgenic silver Yang has stronger photosynthetic capacity, gas exchange capacity and water holding capacity, and with the increase of drought stress, the more obvious the difference. Moderate drought stress of 20 D and 3 under severe drought stress treatment time, the net photosynthetic rate of ABJ01 (Pn) and stomatal conductance (Gs) were significantly higher than 9#; moderate and severe drought stress of 20 D and 30 d, the transpiration rate of ABJ01 (Tr) less than 9# ABJ01, chlorophyll a, chlorophyll b and total chlorophyll content was higher than 9#, severe drought stress reached significant level. Under the condition of moderate and severe drought stress 30 D ABJ01, the Fv/Fm value is higher than 9#, under moderate stress significantly, show that the optical system of ABJ01 damage was smaller than that of 9#.3. and non transgenic poplar compared to JERF36 infiltration in Poplar under drought stress and the ability to regulate the activity of reactive oxygen scavenging capacity. Moderate and severe drought stress 20 d and 30 d ABJ01 (Pro), proline content was higher than 9#, reached a significant level under moderate drought stress. Moderate and severe drought conditions, 20 D and 30 d, superoxide dismutase (SOD) activity in leaves of ABJ01, peroxidase (POD) activity and catalase (CAT) activities were higher than 9# significantly under.4.PEG stress, the transgenic lines had stronger root K~+, Ca~ (2+) absorption capacity under.PEG stress ABJ01 K~+ efflux velocity below 9#, reached a significant level under 10%PEG stress; 20%PEG stress, root tip Ca ABJ01 ~ (2+) flow capacity was significantly higher than that of K element under 9#.20%PEG stress, the root of the ABJ01 was significantly higher than that of 9#, ABJ01 and 20%PEG 10% root stress can accumulate more Ca elements. On the whole, the transgenic poplar under drought stress. The drought resistance obvious growth advantage is mainly reflected in the physiological mechanism by increasing the thickness of palisade tissue reduced, spongy tissue thickness, osmotic adjustment and reactive oxygen scavenging ability, reducing transpiration response to improve its ability to adapt to drought stress, and transgenic poplar under drought stress Pn and gas exchange capacity is effectively improved and the ability to maintain rapid growth. In addition, exogenous JERF36 gene can increase the transgene in silver Yang Genjian cytoplasmic Ca~ (2+) in the flow, reducing the loss of K~+, maintain the apical high Ca~ (2+), K~+ level, to enhance the penetration of transgenic poplar regulation and water absorbing capacity, to enhance gene transfer The tolerance of poplar to drought stress is improved, and the drought resistance of poplar in transgenic silver is improved.

【学位授予单位】：中国林业科学研究院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S792.11

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