本文選題：干旱脅迫 + 轉(zhuǎn)基因銀中楊��； 參考：《中國(guó)林業(yè)科學(xué)研究院》2016年碩士論文

【摘要】：楊樹(shù)(Populus)是世界上分布最廣、栽培面積最大的速生用材林樹(shù)種之一,也是我國(guó)北方地區(qū)最重要的造林樹(shù)種。長(zhǎng)期以來(lái),楊樹(shù)遺傳改良以追求速生性為主,而忽視其抗逆性,致使其在我國(guó)廣大的干旱、半干旱地區(qū)難以發(fā)揮應(yīng)有作用。利用基因工程開(kāi)展楊樹(shù)抗旱品種選育和改良,對(duì)干旱、半干旱荒地利用將產(chǎn)生積極作用。本文以轉(zhuǎn)JERF36基因銀中楊(P.alba×P.berolinensis)(ABJ01)和非轉(zhuǎn)基因銀中楊(9#)為試驗(yàn)材料,通過(guò)測(cè)定土壤控水干旱脅迫條件下植株生長(zhǎng)、葉片解剖結(jié)構(gòu)、光合參數(shù)、生理生化指標(biāo),和水培PEG模擬干旱脅迫條件下根尖離子流等指標(biāo),開(kāi)展了轉(zhuǎn)基因銀中楊的抗旱性評(píng)價(jià)研究,初步揭示轉(zhuǎn)JERF36基因銀中楊抗旱性生理機(jī)制,為進(jìn)一步深入研究轉(zhuǎn)基因植物的抗旱性機(jī)理奠定基礎(chǔ),并為轉(zhuǎn)基因楊樹(shù)的推廣應(yīng)用提供科學(xué)依據(jù)。主要研究結(jié)果如下:1.轉(zhuǎn)JERF36銀中楊在干旱脅迫下生長(zhǎng)優(yōu)勢(shì)明顯,葉片柵欄組織發(fā)達(dá),其抗旱能力得到提高。干旱脅迫處理的3個(gè)時(shí)間點(diǎn)上(10 d、20 d、30 d),ABJ01的苗高、地徑均大于9#,中度干旱脅迫處理20 d時(shí),ABJ01的地徑顯著高于9#,重度干旱脅迫下處理20 d和30 d時(shí),ABJ01的苗高顯著高于9#;中度和重度干旱處理30 d時(shí),ABJ01的單葉面積和總干重顯著大于9#。中度干旱脅迫處理20 d時(shí),ABJ01的葉片厚度顯著高于9#;重度干旱脅迫下,在3個(gè)處理時(shí)間點(diǎn)上ABJ01的葉片上、下表皮厚度均顯著高于9#;中度干旱脅迫處理10 d和30 d時(shí)及重度干旱脅迫處理20 d時(shí),ABJ01柵欄組織厚度顯著高于9#;中度和重度干旱脅迫處理30 d時(shí),ABJ01的海綿組織厚度顯著低于9#。2.干旱脅迫下,轉(zhuǎn)JERF36銀中楊較非轉(zhuǎn)基因銀中楊具有更強(qiáng)的光合能力、氣體交換能力和保水能力,且隨著干旱脅迫程度的增加,差異越明顯。中度干旱脅迫處理20 d和重度干旱脅迫下3個(gè)處理時(shí)間點(diǎn)上,ABJ01的凈光合速率(Pn)和氣孔導(dǎo)度(Gs)均顯著高于9#;中度和重度干旱脅迫處理20 d和30 d時(shí),ABJ01的蒸騰速率(Tr)低于9#,ABJ01的葉綠素a、葉綠素b和總?cè)~綠素含量較9#高,重度干旱脅迫條件下達(dá)到顯著水平。中度和重度干旱脅迫處理30 d時(shí),ABJ01的Fv/Fm值較9#高,中度脅迫條件下達(dá)顯著水平,表明ABJ01的光系統(tǒng)受損情況較9#小。3.與非轉(zhuǎn)基因銀中楊相比,轉(zhuǎn)JERF36銀中楊在干旱脅迫下的滲透調(diào)節(jié)能力和活性氧清除能力更強(qiáng)。中度和重度干旱脅迫處理20 d和30 d時(shí),ABJ01的脯氨酸(Pro)含量均比9#高,中度干旱脅迫下達(dá)到顯著水平。中度和重度干旱脅迫條件下,處理20 d和30 d時(shí),ABJ01葉片的超氧化物歧化酶(SOD)活性、過(guò)氧化物酶(POD)活性及過(guò)氧化氫酶(CAT)活性均高于9#,差異均達(dá)顯著水平。4.PEG脅迫條件下,轉(zhuǎn)基因株系根系具有更強(qiáng)的K~+、Ca~(2+)吸收能力。PEG脅迫下ABJ01的K~+外排流速低于9#,10%PEG脅迫下達(dá)到顯著水平;20%PEG脅迫下,ABJ01的根尖Ca~(2+)內(nèi)流能力顯著高于9#。20%PEG脅迫下ABJ01的根中K元素顯著高于9#,10%和20%PEG脅迫下ABJ01的根系能夠積累更多的Ca元素。總體上,轉(zhuǎn)基因銀中楊在干旱脅迫下生長(zhǎng)優(yōu)勢(shì)明顯。其抗旱生理機(jī)制主要體現(xiàn)在通過(guò)增加?xùn)艡诮M織厚度、降低海綿組織厚度、提高滲透調(diào)節(jié)和活性氧清除能力,減少蒸騰等響應(yīng)來(lái)提高其干旱脅迫適應(yīng)能力,同時(shí)轉(zhuǎn)基因銀中楊在干旱脅迫下Pn以及氣體交換能力得到有效提高,維持快速生長(zhǎng)的能力。另外,外源基因JERF36的導(dǎo)入能夠增加轉(zhuǎn)基因銀中楊根尖胞質(zhì)Ca~(2+)內(nèi)流,減少K~+的損失,維持根尖較高的Ca~(2+)、K~+水平,以增強(qiáng)轉(zhuǎn)基因銀中楊的滲透調(diào)節(jié)能力和吸水能力,來(lái)增強(qiáng)轉(zhuǎn)基因銀中楊的對(duì)干旱脅迫的耐受能力,從而提高轉(zhuǎn)基因銀中楊的抗旱能力。
[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.

【學(xué)位授予單位】：中國(guó)林業(yè)科學(xué)研究院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：S792.11

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