盐胁迫下外源NO对苜蓿幼苗生长及氮代谢的影响

为探寻增强苜蓿耐盐能力的调控途径,以甘农4号苜蓿品种为材料,采用NO供体硝普钠、NO清除剂c-PTIO及硝普钠类似物亚铁氰化钠处理苜蓿幼苗,研究盐胁迫下外源NO对苜蓿幼苗生长、光合特征、氮同化酶活性和氮代谢物含量的影响.结果表明: 外源NO能明显缓解盐胁迫对苜蓿幼苗生长及光合作用的抑制,单株干质量、叶绿素含量、净光合速率、蒸腾速率和可溶性蛋白含量增加;外源NO能增强硝酸还原酶、谷氨酰胺合成酶和谷氨酸合酶活性,抑制蛋白水解酶和谷氨酸脱氢酶活性, 降低叶片中游离氨基酸含量,提高硝态氮含量,加快铵的同化.NO供体SNP的类似物亚铁氰化钠对盐胁迫下苜蓿幼苗生长及氮代谢无调控作用;施用NO清除剂c-PTIO加剧了盐胁迫对苜蓿幼苗生长和氮代谢的抑制,添加外源NO能缓解c-PTIO的抑制效应.盐胁迫下,外源NO和内源NO均参与了苜蓿幼苗氮代谢的调控.     

叶施NO对NaCl胁迫下红砂幼苗叶片和根系中氮代谢酶及营养物质的影响

以当年生红砂(Reaumuria soongorica)幼苗为材料,采用盆栽实验,考察叶面喷施不同浓度(0、0.01、0.10、0.25、0.50、1.00 mmol·L^-1)NO供体硝普钠 (SNP) 对NaCl(300 mmol·L^-1)胁迫下红砂根、叶中可溶性蛋白、游离氨基酸和硝态氮含量,以及谷氨酰胺合成酶(GS)、谷氨酸合酶(GOGAT)、硝酸还原酶(NR)活性的影响,并采用主成分分析和隶属函数法筛选NO对NaCl胁迫缓解效应的氮代谢指标和最佳NO浓度,以探讨外源NO对NaCl 胁迫下红砂缓解效应的氮代谢响应机制。结果表明：(1)在300 mmol·L^-1 NaCl胁迫处理下,红砂幼苗根、叶中可溶性蛋白、硝态氮含量以及GS、GOGAT、NR活性均比对照显著下降。(2)外源NO能显著提高盐胁迫下红砂叶、根中GS、GOGAT、NR活性和硝态氮含量,增加根中可溶性蛋白和游离氨基酸含量。(3)NR和GOGAT活性可用于评价NO对NaCl胁迫下红砂幼苗的缓解作用,外源NO(SNP)对红砂幼苗在NaCl胁迫下的缓解效果强弱表现为0.25 mmol·L^-1> 0.50 mmol·L^-1> 0.10 mmol·L^-1> 1.00 mmol·L^-1> 0.01 mmol·L^-1。研究发现,300 mmol·L^-1 NaCl胁迫显著抑制了红砂幼苗氮代谢,外源NO(SNP)有助于提高盐胁迫下红砂NR活性,加快硝态氮转化为铵态氮,促进红砂叶片和根中GS/GOGAT对转化物的同化,从而增强红砂幼苗的耐盐性,并以0.25 mmol·L^-1SNP处理时缓解作用最佳;NR和GOGAT活性可作为NO缓解盐胁迫的评价指标。     

蒺藜苜蓿MtbHLH148转录因子的克隆与转化及其功能分析

bHLH(Basic helix loop helix, bHLH)转录因子家族是植物最大的转录因子家族之一,广泛参与植物生长发育和盐胁迫应答机制。该研究利用同源克隆的方法克隆蒺藜苜蓿(Medicago truncatula)的MtbHLH148基因,采用qRT PCR方法分析MtbHLH148基因在蒺藜苜蓿中的表达特性,构建超表达载体并通过农杆菌侵染法转化拟南芥(Arabidopsis thaliana),对转基因拟南芥的耐盐性相关功能进行分析研究。结果显示：(1)从蒺藜苜蓿中获得MtbHLH148基因,该基因cDNA全长1 343 bp,包含开放阅读框为603 bp,编码 201 个氨基酸,蛋白分子量22.7 kD,等电点为11.76;蛋白结构分析显示,该蛋白无跨膜结构域,无信号肽,为亲水性蛋白;含有精氨酸／赖氨酸残基的保守结构域和典型的bHLH结构域;二级结构以α 螺旋和无规则卷曲为主。(2)亚细胞定位表明,MtbHLH148蛋白定位在细胞核。(3)进化树分析表明,MtbHLH148与大豆(Glycine max)的亲缘性最近;启动子分析发现,该基因启动子区域含有光响应元件、MYB结合位点以及ABA应答元件ABRE,可能参与非生物胁迫。(4)qRT PCR分析发现,MtbHLH148基因在蒺藜苜蓿的茎中表达量最高,叶中表达量最低,且MtbHLH148基因受ABA(100 μmol/L)诱导并在盐胁迫(200 mmol/L NaCl)处理8 h内表达量上调,而在低温(4 ℃)处理时表达量明显下调。(5)成功构建超表达载体pCAMBIA3301 MtbHLH148并转化拟南芥获得16个抗性株系,经鉴定有12个过表达株系,其中表达量最高的转基因株系为OE8;对OE8株系耐盐性功能分析发现,转基因拟南芥植株的发芽率明显高于野生型,盐胁迫下转基因拟南芥的根长是野生型的1.5倍,表明其耐盐性得到了增强。研究表明,MtbHLH148基因可能在盐胁迫调节机制中具有一定的调控作用。     

氮代谢参与植物逆境抵抗的作用机理研究进展

近年来,植物所受到的诸如干旱、盐、高温、低氧、重金属胁迫和营养元素缺乏等环境胁迫越来越多,严重影响了植物的生长发育及作物的质量和产量。氮素是植物生长发育所需的必需营养元素,同时也是核酸、蛋白质和叶绿素的重要组成成分,其代谢过程与植物抵抗逆境的能力息息相关。氮代谢是指植物对氮素的吸收、同化和利用的全过程,是植物体内基础代谢途径之一。氮代谢主要从氮素吸收、同化及氨基酸代谢等方面参与植物的抗逆性,并通过调节离子吸收和转运、稳定细胞形态和蛋白质结构、维持激素平衡和细胞代谢水平、减少体内活性氧(reactive oxygen species,ROS)生成以及促进叶绿素合成等生理机制来影响植物抵抗非生物胁迫的能力。因此,提高植物在逆境下的氮代谢水平是减轻外界胁迫对其损伤的一种潜在途径。该文从氮素同化的基本途径出发,分别阐述了氮代谢在干旱胁迫、盐胁迫和高温胁迫等多个方面的逆境抵抗过程中的作用机理,为氮代谢参与植物抗逆性研究提供了有利参考。     

NO3-胁迫对草莓幼苗光合特性和氮代谢的影响

沙培条件下,以16 mmol NO₃^-·L^-1为对照,研究不同浓度NO₃^-(64、112 和160 mmol·L^-1)对草莓幼苗光合特性和氮代谢的影响.结果表明： 处理8 d后,随NO₃^-浓度的增加,草莓叶片净光合速率(P_n)、气孔导度(g_s)、PSⅡ实际光化学效率(Φ_PSⅡ)、PSⅡ最大光化学效率(F_v/F_m)和光化学猝灭系数(q_P)均显著降低,当NO₃^-浓度达到160 mmol·L^-1时,较对照分别降低67.7%、68.4%、35.7%、23.2%、26.9%;非光化学猝灭系数(q_N)逐渐升高,64、112 和160 mmol NO₃^-·L^-1处理较对照分别升高4.4%、10.9%、75.8%;胞间CO₂浓度(C_i)呈先降低后升高趋势,气孔限制值(L_s)呈先升高后降低趋势.随NO₃^-浓度增加,草莓叶片及根系中硝态氮、铵态氮、全氮和凯氏氮含量逐渐增加,蛋白氮含量减少.硝酸还原酶(NR)、谷氨酰胺合成酶(GS)、谷氨酸合成酶(GOGAT)、谷氨酸脱氢酶(GDH)活性均随NO₃^-浓度增加呈现先升高后降低趋势.随NO₃^-处理浓度增加草莓幼苗叶片净光合速率下降,PSⅡ电子传递受阻,氮素积累,高浓度下氮代谢酶活性降低,营养液中NO₃^-浓度为64 mmol·L^-1时开始产生胁迫,不利于草莓幼苗的生长.     

水分胁迫对荔枝叶片氮和核酸代谢的影响及其与抗旱性的关系

水分胁迫下,荔枝叶片蛋白酶活性和Ｐｒｏ含量增加,ＰＤＨ活性与可溶性蛋白质含量下降,抗旱性强的品种蛋白酶活性增加的幅度蛋白质含量下降的幅度小于抗旱性弱的品种,而ＰＤＨ活性下降的幅度和Ｐｒｏ含量上升的幅度均大于抗旱性弱的品种。水分胁迫引起荔枝叶片核酸,ＤＮＡ和ＲＮＡ含量下降,ＤＮＡ含量下降的幅度小于ＲＮＡ含量下降的幅度;ＤＮａｓｅ和ＲＮａｓｅ活性上升,ＤＮａｓｅ活性上升的幅度小于ＲＮａｓｅ活性上升的幅     

ABA对盐胁迫下侧柏活性氧代谢及其相关基因表达的研究

以侧柏幼苗为试验材料,采用水培方式,研究不同浓度NaCl(0、100、200、300和400mmol·L~(-1))以及ABA(0、0.5、1、10、100和200μmol·L~(-1)处理对盐胁迫下侧柏幼苗活性氧代谢的影响。结果显示:(1)随着NaCl处理浓度增加,侧柏幼苗叶片过氧化氢(H_2O_2)、丙二醛(MDA)、谷胱甘肽(GSH)和脯氨酸含量,以及抗氧化物酶(SOD、POD和CAT)活性呈上升趋势,而可溶性蛋白含量降低。(2)于300mmol·L~(-1)NaCl胁迫处理48h后进行ABA(1和10μmol·L~(-1))处理,分析发现侧柏幼苗叶片的SOD、POD和CAT活性提高,GSH、脯氨酸和可溶性蛋白含量增加,H_2O_2和MDA的积累减少。(3)NaCl胁迫后,侧柏幼苗叶片活性氧代谢相关基因(Cu/Zn-SOD、CAT、GR、APX、MDAR和GST)的表达水平随ABA浓度增加呈先升后降的趋势,并于10μmol·L~(-1) ABA处理48h时达到峰值。研究表明,适宜浓度ABA通过促进盐胁迫下侧柏幼苗叶片的抗氧化物酶活性,提高渗透调节能力,增加GSH含量,降低H_2O_2和MDA积累,从而有效降低活性氧对侧柏叶片的伤害,保护细胞膜结构的完整,增强其抗盐性。     

水分胁迫对荔枝叶片氮和核酸代谢的影响及其与抗旱性的关系

水分胁迫下,荔枝叶片蛋白酶活性和Ｐｒｏ 含量增加,ＰＤＨ 活性和可溶性蛋白质含量下降;抗旱性强的品种蛋白酶活性增加的幅度和蛋白质含量下降的幅度小于抗旱性弱的品种, 而ＰＤＨ 活性下降的幅度和Ｐｒｏ 含量上升的幅度均大于抗旱性弱的品种。水分胁迫引起荔枝叶片核酸、ＤＮＡ 和ＲＮＡ 含量下降,ＤＮＡ 含量下降的幅度小于ＲＮＡ 含量下降的幅度;ＤＮａｓｅ 和ＲＮａｓｅ 活性上升,ＤＮａｓｅ 活性上升的幅度小于ＲＮａｓｅ 活性上升的幅度。抗旱性强的品种核酸、ＤＮＡ 和ＲＮＡ 含量下降的幅度以及ＤＮａｓｅ 和ＲＮａｓｅ 活性上升的幅度均小于抗旱性弱的品种。     

溶藻细菌对固氮藻氮代谢的影响

随着水体富营养化的加剧,有害藻类水华和赤潮的爆发日趋频繁,导致水质的恶化,水体资源丧失其功能和价值。修复藻型富营养化水体生态系统可以从控制藻类繁殖和调整藻类群落结构方面着手,使整个水生     

重庆市主城区不同生境类型的蝴蝶多样性

2006年9-11月,选取重庆主城区人工次生林、溪流沿岸农田、山地农田、天然次生林和人工园林等5种不同的生境类型,进行蝴蝶调查.共采集蝴蝶529只,隶属于6科16属20种.研究区优势种蝶类为酢浆灰蝶(Pseudozizeeria maha)(相对多度为38.75%)和菜粉蝶(Pieris rapae)(相对多度为17.01%).5种生境类型的多样性指数变化趋势为天然次生林>溪流沿岸农田>山地农田>人工次生林>人工园林.同时分析了人为干扰和生境类型与蝴蝶多样性的关系.结果表明,人为干扰对蝴蝶多样性的负面影响尤其严重,保留城区内残存的农田生境斑块对蝴蝶多样性的保护有积极的意义.     

联合固氮的研究进展

1975年,Dbereiner实验发现与禾本科植物联合共生的固氮菌并提出根际联合固氮的概念。近年来,随着一些新的研究手段包括化学分析、遗传工程、分子生物学、免疫学等方法的运用,推进了联合固氮领域的研究深度。综述了近年来发现的联合固氮菌的种类;联合固氮体系的形成过程（趋化、结合和侵入）;影响联合固氮的主要因素：自然条件、土著微生物的竞争、植物基因型差异和环境条件的变化、结合态氮（氨、亚硝酸盐、硝酸盐等）和氧。并从固氮联合作用测定方法、联合固氮菌的资源筛选、联合固氮工程菌的研究、联合固氮分子生态学研究方法4个方面,论述了联合固氮的研究现状。     

联合固氮菌的合成生物学研究进展

氮素是作物生长过程中最重要的元素,氮素缺乏将会严重影响作物生长。随着人类对粮食的需求量增加,化学氮肥的施用量越来越多。生物固氮在全球氮素循环中有着重要的作用,60%的氮来源于生物固氮。因此,生物固氮,尤其是能够在作物中定殖的联合固氮菌,最有可能代替氮肥成为粮食作物的主要氮源。长期以来,如何提高生物固氮效率以及在作物中实现生物固氮是生物学家的重要研究方向。合成生物学的出现和发展为能够生物固氮的研究带了新的机遇,有望缓解粮食作物对化学氮肥的大量需求。本文概述了固氮菌的种类、联合固氮菌中固氮基因岛的组成以及转录调控机理,阐述了合成生物学在生物固氮领域中的研究现状,对未来的联合固氮菌合成生物学的发展方向作出了展望。     

Nitrogen-fixing Bacillus sp. associated with Douglas-fir tuberculate ectomycorrhizae

Nitrogenase activities, measured by acetylene reduction, were detected under microaerophilic field conditions in Douglas-fir tuberculate ectomycorrhizae. Tuberculate ectomycorrhizae consist of densely packed clusters of ectomycorrhizal rootlets enclosed in a supplementary fungal peridium-like layer. Nitrogenase activity was primarily in the external layer and was greatly enhanced with added sucrose. The bacterium isolated, a nitrogen-fixing, spore-forming Bacillus sp., is an aerobe but requires anaerobic conditions for nitrogenase activity. Respiration in the tuberculate complex by the fungus, roots, and associated mycorrhizosphere microbes probably contributes to maintaining a microaerophilic niche where nitrogen fixation can take place. Water extracts of peridium or mycorrhizal root tips enhanced nitrogenase activity of this associative Bacillus sp., thereby indicating a close nutritional relationship between this bacterium and the tuberculate mycorrhizae. Thiamine more significantly enhanced bacterial nitrogenase activity than biotin; no activity was detected with p-aminobenzoic acid. Even though the levels ofnitrogenase activities in the tubercles in situ were low, as measured by the present methods, they may indicate a significant contribution to the nitrogen dynamics of these nitrogen-limited Douglas-fir forests over a long-term period.     

植物根压研究进展

根压是植物根部产生的一种静水压力,广泛存在于多种植物中。在蒸腾作用很弱的情况下,根压不但可驱动水分从根部流向冠层叶片,缓解因白天强烈蒸腾而导致的水分亏缺,而且在木质部导管栓塞修复方面具有重要作用。虽然国内外学者对根压的产生已有一些解释,普遍接受的观点有渗透理论、代谢理论和水分向上共同运输假说等,但根压产生的机制至今仍是科学家争议的焦点之一。根压的测定方法虽有直接和间接测定、损伤和无损伤测定之分,但较为先进的根压测定技术仍需进一步改善和提升。受水通道蛋白、遗传因素、生境等因素的影响,根压的大小存在差异,即使是较低的根压也会影响农作物生长。在促进转运蛋白质、酶、氨基酸、激素及钙元素等在农作物木质部和韧皮部之间流通方面,适当大小的根压发挥重要作用,且有助于提高农作物产量。因此,加深对植物根压的认识和理解具有重要的生物学意义。该文从根压的定义和产生机制、具有根压的植物类群、根压的测定方法和大小、影响根压的主要因素及根压在植物科学研究领域的意义和影响等多个方面分别进行了归纳总结,并结合当前研究热点和研究成果,针对植物根压研究过程中遇到的问题和后续研究趋势及方向进行了展望。     

Inducing drought tolerance in plants: Recent advances

Undoubtedly, drought is one of the prime abiotic stresses in the world. Crop yield losses due to drought stress are considerable. Although a variety of approaches have been used to alleviate the problem of drought, plant breeding, either conventional breeding or genetic engineering, seems to be an efficient and economic means of tailoring crops to enable them to grow successfully in drought-prone environments. During the last century, although plant breeders have made ample progress through conventional breeding in developing drought tolerant lines/cultivars of some selected crops, the approach is, in fact, highly time-consuming and labor- and cost-intensive. Alternatively, marker-assisted breeding (MAB) is a more efficient approach, which identifies the usefulness of thousands of genomic regions of a crop under stress conditions, which was, in reality, previously not possible. Quantitative trait loci (QTL) for drought tolerance have been identified for a variety of traits in different crops. With the development of comprehensive molecular linkage maps, marker-assisted selection procedures have led to pyramiding desirable traits to achieve improvements in crop drought tolerance. However, the accuracy and preciseness in QTL identification are problematic. Furthermore, significant genetic × environment interaction, large number of genes encoding yield, and use of wrong mapping populations, have all harmed programs involved in mapping of QTL for high growth and yield under water limited conditions. Under such circumstances, a transgenic approach to the problem seems more convincing and practicable, and it is being pursued vigorously to improve qualitative and quantitative traits including tolerance to biotic and abiotic stresses in different crops. Rapid advance in knowledge on genomics and proteomics will certainly be beneficial to fine-tune the molecular breeding and transformation approaches so as to achieve a significant progress in crop improvement in future. Knowledge of gene regulation and signal transduction to generate drought tolerant crop cultivars/lines has been discussed in the present review. In addition, the advantages and disadvantages as well as future prospects of each breeding approach have also been discussed.     

Nitrogen fixation associated with grasses and cereals: Recent results and perspectives for future research

Over the last few years research in the area of biological nitrogen fixation (BNF) associated with cereals and grasses has become divided into two areas. On the one hand there have been a large number of reports of responses of field-grown plants to inoculation with N₂-fixing bacteria, principallyAzospirillum spp. On the other hand there have been several reports of significant contributions of associated BNF to the nutrition of several crops, including wetland rice, sugar cane and some forage grasses. However, where BNF contributions have definitely been established no certain information is available as to the diazotrophic organisms responsible. Furthermore, certain recent reports indicate that, at least in some cases, responses of plants to inoculation withAzospirillum spp. have been shown not to be due to BNF contributions. In this paper we review some recent progress in this field, particularly at our institute in Rio de Janeiro, concerning specificity of selected Azospirillum strains in the infection of cereal roots and the promotion of responses in the host plants. The possible mechanisms of plant response are discussed including the possibility that plant growth substances or bacterial nitrate reductase are involved. The application of¹⁵N and N balance techniques to the quantification of plant associated BNF are considered and the possible strategies that may be adopted to further the understanding of true N₂-fixing plant/diazotroph associations. The recent discovery of many more plant-associated N₂-fixing bacteria suggests that further research in this area may eventually lead to the development of such associations with applications for agricultural productivity.     

Sensitivity of selected Frankia isolates from Casuarina, Allocasuarina and North American host plants to sodium chloride

Three experiments examined the effects of NaCl concentrations 0 to 500 mM on the growth of isolates of Frankia from Casuarinaceae and selected North American host plants. Four Casuarina isolates grew well in defined medium (pyruvate-BAP) but not in a yeast extract medium. Conversely the non-Casuarina isolates preferred the yeast-extract medium, although two of them grew in the defined medium. When grown in their preferred medium, the Casuarina isolates were little affected by NaCl concentrations up to 200 m M but did not grow at 500 m M . The non-Casuarina isolates, with the exception of an isolate from Purshia tridentata . were severely affected above 50 m M NaCl.
Nitrogenase activity (C₂H₂ reduction) by the non-Casuarina isolates could not be detected in low-N medium although protein determinations indicated that a low level of nitrogen fixation had occurred. All four Casuarina isolates showed nitrogenase activity in culture, up to 200 m M NaCl, although at that concentration of NaCl, growth was affected more than that of cultures in N-supplemented medium. All four strains showed a marked increase in nitrogenase activity up to 72 h after the addition of C₂H₂, with the magnitude of the effect and their subsequent behaviour being strain dependent.
The results indicate that the isolates of Frankia from Casuarina and Allocasuarina , and that from Purshia tridentata , are more tolerant of NaCl than isolates from species not normally growing under sodic conditions. They provide optimism that these strains could successfully establish in saline soils if introduced with species of host plants tolerant to these soils.