【作者】 张磊；

【导师】 张含国；

【作者基本信息】 东北林业大学 ， 林木遗传育种， 2012， 博士

【摘要】 长白落叶松(Larix olgensis Henry)是我国北方主要速生针叶用材树种,在我国东北地区的造林面积所占比重较大。本研究对22个4年生长白落叶松种内控制授粉家系在干旱胁迫条件下的生长、光合、生理和差异表达蛋白进行研究。每15天进行一次测量,共四次。对苗高、净光合速率、蒸腾速率、气孔导度、MDA、POD、可溶性蛋白和叶绿素共8个指标进行了综合分析。同时对胁迫第30天的长白落叶松针叶样品和对照进行旱胁迫下差异蛋白表达的研究研究表明：长白落叶松家系间存在着丰富的变异,在生长、光合、生理等各项指标中家系间差异显著,有利于优良抗旱家系的选择。不同家系在不同指标的表现能力有所不同,说明每个家系在干旱胁迫的条件下以不同的新陈代谢反应来抵抗外界环境造成的影响。长白落叶松家系内也存在着丰富的变异。家系间气孔导度变异系数最高为103.79%,不同家系间气孔导度的变异系数范围为28.72%-76.64%；叶绿素含量变异系数最低为12.86%,不同家系的叶绿素含量的变异系数范围为4.8%-27.5%,差异最小。POD不同家系的变异系数从40.3%-130.8%,差异最大。家系2×1不同指标的变异系数从10.8%(苗高)-130.8%(POD含量),变化幅度最大家系34×1的苗高生长旺盛.比最低的2×2高出57.8%.比家系平均值高出18.5%。14x3,34x1,1×14,14x34家系在整个胁迫过程中苗高一直高于对照,尤其是在胁迫后期表现稳定,4个家系在旱胁迫末期的苗高平均值高于所有家系苗高平均值17.5%,高于对照3.7%,高于在整个旱胁迫期间均生长较差的4个家系在胁迫末期苗高平均值50.09%。胁迫开始前生长最好的家系比最差的家系高出132.06%,高于对照35.49%,胁迫末期最好的家系比最差的家系高出135.91%,高于对照仅0.25%。说明即使是苗高生长表现优良的家系,在干旱胁迫的条件下也受到了很大程度的影响。家系母本为73-14的情况最好,比母本为73-3的苗高高出28.18%,比平均值高出10.32%与对照相比,净光合速率、气孔导度、蒸腾速率、POD叶绿素在干旱胁迫的条件下呈下降趋势,MDA显著增加,可溶性蛋白先增加后降低。且不同家系在对于旱胁迫的响应出现在胁迫前期、中期或者后期,有所不同。苗高,净光合速率,气孔导度,POD含量在母本不同、父本不同的家系间均差异显著,MDA含量,蒸腾速率,可溶性蛋白在父本不同的家系间差异显著,叶绿素含量在母本不同的家系间差异显著。在长白落叶松家系干旱胁迫和对照的样品中提取蛋白质组,经过双向电泳分析后找到23个差异表达的蛋白点,最终鉴定了18个差异蛋白点。根据生物学功能,将这18个鉴定蛋白质分为信号转导相关蛋白、ATP相关酶、光合调控相关蛋白、代谢相关蛋白、降解有害物质相关酶、其他蛋白六类。苗高与各项指标的相关性均不显著。净光合速率与苗高为正相关,但是相关不显著,与POD和蒸腾速率均为极显著正相关。MDA与可溶性蛋白为显著正相关,与其他指标为负相关,其中与POD,净光合速率和蒸腾速率都达到显著负相关水平。通过隶属函数综合评定各项指标,长白落叶松22个家系的抗旱能力各有不同。34×2,2×1和34×34家系的抗旱能力最强,2×2,3×1和3×14家系的抗旱能力最差。第1名34×2家系比最后1名2×2的隶属函数平均值高出32.49%,比平均值高出10.99%。其中苗高占比例最大为18.37%,在隶属函数中为最可靠鉴定指标。更多还原

【Abstract】 Larix olgensis Henry is one of the most important conifer species in China. It is a fast-growing timber species and shares the largest afforestation area in northeast China. The study take22L. olgensis control pollinated families of4-years-old as the materials and the experiment materials were divided into control group and drought stress treatment group, then studied on the growth, photosynthesis, physiology and drought stress-responsive protein expression, finally made a comprehensive evaluation of height, photosynthesis, transpiration, MDA, POD, soluble protein and chlorophyll. And the conifer of day-30and control were also used for research of differentially expressed proteins.The results showed that there were rich variations among the families and family’s variation was significant difference in height, photosynthesis and physiology indicator, it was helpful for the excellent family selection. Each family had a different performance under the drought stress for its self-protection mechanism is different.There were rich variations within the families, too. The most variation coefficient was the one of stomatal conductance and it was103.79%, the range of families were from28.72%to76.64%; the least variation coefficient was the one of chlorophyll and it was12.86%, the range of families were from4.8%to27.5%which was the smallest difference of families. The POD variation coefficient range of families was from40.3%to130.8which was the most difference among families. The variation coefficient range of family2×1was the most, it was from10.8%(height) to130.8%(POD).Among all families, family34×1’s growth was significantly higher than the control and other families; it was57.8%more than the slowest growth family2×2and18.5%more than the average. The height of family14×3,34×1,1×14,14×34was all more than the control during the whole drought period, the height average of these4families were17.5%more than the average of all families.3.7%more than the control.50.09%more than the one of4worst families. On day0. the height of the best family was132.06%more than the worst one,35.49%more than the control, while on day45. the height of best family was135.91%more than the worst one, only0.25%more than the control. It indicated that even the better families’ height was decrease under drought stress.Compared with the control, photosynthesis rate, stomatal conductance, transpiration rate, POD, chlorophyll were downward trend under drought stress while height increased slowly, MDA increased significant, soluble protein first increased and then decreased. The performance of families was different during the different drought period. Height, photosynthesis rate, stomatal conductance, POD was significant difference between families of different female or male parent; MDA, transpiration rate and soluble protein was significant difference between families of male parent, while chlorophyll was significant difference between families of female parent.Proteins were extraction from the control and drought stress samples,23differentially expressed proteins were found and18were indentified after analysis. According to biological function, these drought stress-responsive protein were classified into6groups:protein related to transduction, enzyme related to ATP synthesis, protein related to photosynthetic regulation, protein related to metabolism, enzyme to degradation hazardous substance, others.Height has no significant correlation with other index. Photosynthesis was positive correlation with height but no significant, significant positive correlation with POD and transpiration rate. MDA was significant positive correlation with soluble protein while negative correlation with other index, of which POD, photosynthesis rate and transpiration rate was significant.22L. olgensis families had different drought resistance capacity according to the membership function.34×2,2×land34×34are the best3families to drought stress,2×2,3×1and3×14were the worst3ones. Height was the most reliable index of all. The first one34×2was32.49%than the worst one2×2. and10.99%more than the average. Height had the most proportion (18.37%) among all index of membership function. so it was the most reliable indicator to identification of drought resistance.更多还原