AtSOS基因在紫花苜蓿中的表达及其耐盐性研究

本研究采用基因工程技术改良紫花苜蓿耐盐性,通过种植转基因耐盐紫花苜蓿达到改良土壤的目的。以紫花苜蓿子叶节为外植体,通过农杆菌介导法将来源于拟南芥的AtSOS1-AtSOS2-AtSOS3多基因表达载体导入阿尔冈金紫花苜蓿中,经PCR检测、抗除草剂筛选和RT-PCR鉴定,获得了能稳定表达的转基因株系。以转基因的紫花苜蓿和野生型紫花苜蓿为材料进行盐处理,每个处理重复3次,测定其生理生化指标、株高、Na⁺和K⁺含量、细胞膜透性、叶绿素含量。结果显示,在不同盐浓度处理下,所有植株的株高均有所增长,但在100和200 mmol·L^-1的NaCl处理下,转基因植株的长势显著高于野生型植株;随着处理时间的增加,所有植株的叶绿素含量均呈先上升后下降的趋势,且野生型植株叶绿素含量均低于转基因植株;在100和200 mmol·L^-1的NaCl处理下,转基因植株的细胞膜透性、超氧化物歧化酶活性和脯氨酸含量的增加量均小于野生型植株,而过氧化物酶、过氧化氢酶活性和可溶性糖含量的增加量均大于野生型;各植株中丙二醛含量均下降,且野生型植株下降的更为明显;盐处理后,转基因植株根系中Na⁺的积累比野生型植株少,而K⁺的吸收多于野生型植株。转AtSOS基因的紫花苜蓿通过发挥AtSOS途径的作用,促进了植物体将细胞内的Na⁺外排,从而减轻盐胁迫对植物体的伤害,提高了转基因植株的耐盐性。     

铺地竹叶片对结球生菜种子发芽及幼苗生长的影响

采用基质盆栽试验,研究了铺地竹(Sasa argenteastriatus E.G.Camus)叶片对结球生菜(Lactucasativa L.var.capitata L.)种子萌发和幼苗生长的影响,结果表明,铺地竹叶片粉末对结球生菜种子的萌发和幼根生长具有一定的抑制作用,随着浓度的增加抑制作用增强,在50～200 g/m2的范围内,幼根抑制率为12.15%～28.91%。在50～100 g/m2的用量下,铺地竹叶片对结球生菜苗长表现为促进生长作用,而在200 g/m2时,则表现为抑制生长作用。施用铺地竹叶片后3 d、6 d、9 d和12 d,结球生菜幼苗的叶绿素含量特别是叶绿素a的含量显著减少,随着基质中叶片粉末浓度的增加,结球生菜叶绿素含量降低,叶绿素a含量高低依次为对照、低浓度处理、中浓度处理、高浓度处理。随着处理后时间的延长,竹叶对结球生菜叶片叶绿素a含量的影响减弱。这个结果进一步验证了自然条件下铺地竹叶片对植物生长的化感效应,说明其具有开发植物源除草剂的潜力,为研究铺地竹叶片中的化感物质提供了佐证。     

Copper‐Induced Oxidative Stress and Responses of the Antioxidant System in Roots of Medicago sativa

The effects of various copper (Cu) concentrations on the antioxidative system in the roots of Medicago sativa were explored. The results indicated that the Cu content of the roots reached a value of 854 μg g^?1 DW at 10 μm Cu and a value of 4415 μg g^?1 DW at 100 μm Cu, suggesting that M. sativa has better ability to tolerate and accumulate Cu than other Cu‐bioaccumulators, and is a potential plant for phytoremediation. Treatment with Cu resulted in a significant increment in the levels of H₂O₂, O₂˙^? and OH˙. The reduced form of ascorbate and glutathione reached a peak at 30 μm Cu, and was followed by a sharp depletion to a lower level than that of the control. In contrast, the levels of the oxidised forms of ascorbate and glutathione showed a progressive increment with increasing Cu concentrations, suggesting that the antioxidant system was unable to cope with Cu stress at higher Cu levels. Under the Cu concentrations tested, the activity of catalase (CAT, EC 1.11.1.6), ascorbate peroxidase (APX, EC 1.11.1.11) and glutathione reductase (GR, EC 1.6.4.2) increased at lower Cu concentrations, and then decreased, reaching a maximum at 30 μm of Cu for APX and GR, at 10 μm for CAT, whereas the activities of guaiacol peroxidase (POD, EC 1.11.1.7) were gradually increased with increasing Cu concentrations. PAGE analysis of superoxide dismutase (SOD, EC 1.1.5.1.1) revealed that one band is a Mn‐SOD and five bands are identified as Cu, Zn‐SOD, whereas Fe‐SOD isoforms were not found in the roots of alfalfa. Cu at 10–100 μm increased the intensity of constitutive isozymes of CAT, APX and POD, whereas it decreased the intensity of isozymes of glucose‐6‐phosphate dehydrogenase (G6PDH, EC 1.1.1.49) significantly. The activities of lipoxygenases (LOX, EC 1.13.11.12) were gradually augmented with increasing Cu concentrations, demonstrating that LOXs are probably involved in production of lipid hydroperoxides and superoxide anion. There was a continuous and pronounced enhancement in the activity of esterase (EST, EC 3.1.1.1) in roots treated with 10–30 Cu μm , whereas EST activity in roots exposed to above 30 μm Cu declined, suggesting that EST plays a protective role under lower Cu concentrations stress.     

水稻根系育种研究进展

水稻（Oryza sativa L.）根系对水稻的生长发育及产量形成具有至关重要的作用。根据目前水稻根系育种的研究现状，分析了水稻根系育种对提高产量、改变形态特征和生理特性的重要性，并对其研究前景进行了分析和展望。     

不同裂荚特性箭筈豌豆裂荚力数字化评价

以美国种质资源库(NPGS)提供的541份箭筈豌豆(Vicia sativa)种质为基础,经过连续3年裂荚率评价,共筛选出高、中、低裂荚种质33份。以这33份箭筈豌豆种质作为本研究的试验材料,利用艾德堡HP-50数显推拉力计结合裂荚率对箭筈豌豆荚果水平放置时的裂荚机械力和垂直放置时的裂荚机械力进行分析,为选育箭筈豌豆抗裂荚种质提供精确、快速的评价方法。结果显示,1)箭筈豌豆高裂荚种质荚果裂荚机械力极显著低于低裂荚种质荚果裂荚机械力(P0.01)。2)箭筈豌豆荚果水平放置时的裂荚机械力极显著低于垂直放置时的裂荚机械力(P0.01)。3)箭筈豌豆荚果在水平放置下裂荚机械力大于9.286N的为低裂荚种质,裂荚机械力小于2.163N的为高裂荚种质。研究表明,利用推拉力计测试水平放置的箭筈豌豆荚果裂荚机械力可准确反映箭筈豌豆不同种质的裂荚特性,并为箭筈豌豆裂荚特性的标准化测定提供了重要的参考依据。     

水稻半外卷叶突变体sol1的表型分析与基因定位

适度卷曲有利于提高水稻叶片的光合效率,增加植株光合产物的有效积累量。我们利用甲基磺酸乙酯(EMS)处理籼型水稻保持系西农1B,获得一个稳定遗传的水稻半外卷叶突变体。该突变体从十叶期开始各叶片逐渐向外卷曲直至半卷状,并伴随茎秆半矮化和叶片披垂,暂被命名为semi-outcurved leaf 1(sol1)。与野生型(WT)相比,sol1的叶片卷曲指数均达到30%以上(P<0.01);倒一、倒二、倒三、倒四节节间长度和穗长极显著缩短,倒一、倒二、倒三叶的叶夹角显著或极显著增加;有效穗数、千粒重、每穗实粒数、结实率显著或极显著下降,一次枝梗数则增加11.3%(P<0.05)。sol1的蒸腾速率、胞间CO2浓度、气孔导度显著高于野生型。石蜡切片显示,sol1倒一叶的泡状细胞体积变小,数量显著增多,表皮细胞体积略微增大。遗传分析表明,sol1的半外卷叶性状受1对隐性核基因调控,定位于6号染色体标记JY6-3和JY6-10之间165 kb的物理范围内,共含15个注释基因。qRT-PCR结果表明,与泡状细胞相关的内卷基因和外卷叶基因RL14、Roc5、REL1在突变体sol1中呈不同程度的上调,NRL、BRD1、OsHox32、ADL1、LC2则呈不同程度的下调。研究结果为SOL1基因的克隆和功能研究奠定了基础。     

有机生态栽培基质配比对生菜生长及品质的影响

以传统基质草炭、蛭石及腐熟羊粪和玉米秸秆为原料,配制成12种不同配比的栽培基质,以商品叶菜类栽培基质作为对照,研究不同基质配方对生菜生长的影响,以期获得最适合生菜栽培的基质配比组合。结果表明,不同组合基质配比均能满足生菜的正常生长,所有处理的株高、叶面积、单株鲜重和单株干重都较对照组(CK)有着不同程度提高,其中T11(草炭∶腐熟羊粪∶玉米秸秆∶蛭石=1∶4∶3∶4)条件下各指标表现最好,分别较CK提高了31.80%、47.15%、59.43%和79.53%;从生理生化指标来看,T11处理的Vc含量最高,是CK的2.5倍;T10(草炭∶腐熟羊粪∶玉米秸秆∶蛭石=3∶1∶4∶4)的根系活力是所有处理中最高的,达到CK的2.02倍;T12(草炭∶腐熟羊粪∶玉米秸秆∶蛭石=3∶4∶1∶4)处理的叶绿素含量最高,是CK的1.41倍,硝酸盐含量最低,较CK降幅达到74.59%。通过隶属函数进行综合比较分析,我们认为T11是所有供试处理中最适合生菜栽培的基质配比。     

Genome‐Wide Identification of MicroRNAs Responsive to High Temperature in Rice (Oryza sativa) by High‐Throughput Deep Sequencing

MicroRNAs (miRNAs) are known to play important roles in plant growth and stress response. Heat stress is a severe abiotic stresses by adversely affecting plant growth and yield. To identify heat‐responsive miRNAs at the genome‐wide level in rice (Oryza sativa), we constructed two small RNA libraries from young panicles treated or not with heat conditions. Ion torrent sequencing of the two libraries identified 294 known miRNAs and 539 novel miRNAs. Differential expression analysis showed that 26 miRNAs were downregulated and 21 miRNAs were upregulated in response to heat stress. Among them, five heat‐responsive miRNAs, including miR162b, miR529a‐p5, PC‐5P‐62245‐9, miR171b and miR169n, were validated by quantitative real‐time polymerase chain reaction. A total of 44 target genes of the differentially expressed miRNAs were predicted. These target genes are most significantly overrepresented in the cell growth process. The results demonstrated that rice miRNAs play critical roles in the heat stress response. This study opens up a new avenue for understanding the regulatory mechanisms of miRNAs involvement in the heat stress response in rice.     

44个紫花苜蓿品种的酸铝适应性与耐受性评价

为了解紫花苜蓿在贵州地区的适应性及耐酸铝胁迫机理，以44份紫花苜蓿品种为研究对象，研究紫花苜蓿处于酸铝胁迫下的生理变化，并揭示其生理变化与耐酸铝胁迫间的关系。利用基因与环境互作模型对两个地点1年的紫花苜蓿进行产量分析，筛选出阿尔冈金、新疆大叶苜蓿、Trifecta、Vernal和中牧1号苜蓿5个耐酸铝强适应品种。利用敏感型UC-1465和耐受型阿尔冈金进行酸铝胁迫试验。结果表明：相同处理下，耐受型紫花苜蓿的电导率、相对铝含量、死亡率显著低于敏感型；紫花苜蓿对酸铝胁迫的响应主要通过柠檬酸、苹果酸、乙酸、酒石酸、反丁烯二酸和草酸的显著（P<0.05）增加来体现，其中苹果酸的合成和分泌增多可能是其耐酸铝胁迫的重要原因。