植物抗盐分子机制及作物遗传改良耐盐性的研究进展

盐胁迫是全球农业生产上的一个主要逆境因子。解析耐盐分子机制有助于培育耐盐能力提高的作物新品种。我们综述了植物对盐胁迫的感应及信号传导、主要Na^＋运输体、盐胁迫下的解毒途径以及耐盐途径中涉及到的表观遗传研究。此外,我们还讨论了利用遗传改良手段提高作物耐盐性的研究进展。     

Physiological and Biochemical Characteristics and Response Patterns of Salinity Stress Responsive Genes (SSRGs) in Wild Quinoa (<i>Chenopodium quinoa</i> L.)

Cultivating salt-tolerant crops is a feasible way to effectively utilize saline-alkali land and solve the problem of underutilization of saline soils. Quinoa, a protein-comprehensive cereal in the plant kingdom, is an exceptional crop in terms of salt stress tolerance level. It seems an excellent model for the exploration of salt-tolerance mechanisms and cultivation of salt-tolerant germplasms. In this study, the seeds and seedlings of the quinoa cultivar Shelly were treated with different concentrations of NaCl solution. The physiological, biochemical characteristics and agronomic traits were investigated, and the response patterns of three salt stress-responsive genes (SSRGs) in quinoa were determined by real-time PCR. The optimum level of stress tolerance of quinoa cultivar Shelly was found in the range of 250–350 mM concentration of NaCl. Salt stress significantly induced expression of superoxide dismutase (SOD), peroxidase (POD), and particularly betaine aldehyde dehydrogenase (BADH). BADH was discovered to be more sensitive to salt stress and played an important role in the salt stress tolerance of quinoa seedlings, particularly at high NaCl concentrations, as it displayed upregulation until 24 h under 100 mM salt treatment. Moreover, it showed upregulation until 12 h under 250 mM salt stress. Taken together, these results suggest that BADH played an essential role in the salt-tolerance mechanism of quinoa. Based on the expression level and prompt response induced by NaCl, we suggest that the BADH can be considered as a molecular marker for screening salt-tolerant quinoa germplasm at the early stages of crop development. Salt treatment at different plant ontogeny or at different concentrations had a significant impact on quinoa growth. Therefore, an appropriate treatment approach needs to be chosen rationally in the process of screening salt-tolerant quinoa germplasm, which is useful to the utilization of saline soils. Our study provides a fundamental information to deepen knowledge of the salt tolerance mechanism of quinoa for the development of salt-tolerant germplasm in crop breeding programs.     

植物根系耐盐机制的研究进展

植物根系能够摄取土壤环境中的养分与水分,在植物的生长发育中起重要的作用。植物根系由于直接与土壤环境相接触会受到非生物胁迫较大的影响。盐胁迫是主要的非生物胁迫之一,对植物根系会产生较大的伤害。综述根系在组织形态和细胞水平上对盐胁迫的应答,以及根系响应盐胁迫的信号传导途径、转录因子与基因,对植物根部耐盐机制的解析和植物耐盐基因工程工具基因的挖掘具有重要意义。     

拟南芥液泡膜Na+/H+逆向转运蛋白研究进展

盐分是植物生长发育的主要限制因素之一,而离子在胞内区室之间的选择性运动对提高植物耐盐性是至关重要的。来自于拟南芥(Arabidopsis thaliana)的AtNHX1基因可编码Na /H 逆向转运蛋白,而Na /H 逆向转运蛋白AtNHX1可将细胞质中多余的Na 排进液泡来消除Na 的毒害,维持细胞的渗透平衡,提高植物的耐盐性。简要综述了AtNHX1基因及Na /H 逆向转运蛋白AtNHX1的特征,AtNHX1的耐盐机制以及植物耐盐基因工程改良等方面的研究进展。     

In vitro and in vivo screening for the identification of salt-tolerant sugarcane (Saccharum officinarum L.) clones: molecular,biochemical, and physiological responses to salt stress

Sugarcane is a glycophyte whose growth and yield can be negatively affected by salt stress. As the arable lands with potential saline soils expand annually, the increase of salt-tolerance in sugarcane cultivars is highly desired. We, herein, employed in vitro and in vivo conditions in order to screen sugarcane plants for salt tolerance at the cellular and at the whole plant levels. Calli of sugarcane cv. Khon Kaen 3 (KK3) were selected after culturing in selective media containing various NaCl concentrations, and regenerated plants were then reselected after culturing in selective media containing higher NaCl concentrations. The surviving plants were finally selected after an exposure to 254 mM NaCl under greenhouse conditions. A total of 11 sugarcane plants survived the selection process. Four plants that exhibited tolerance to the four different salt concentrations applied during the aforementioned screening process were then selected for the undertaking of further molecular, biochemical, and physiological studies. The construction of a dendrogram has revealed that the most salt-tolerant plant was characterized by the lowest genetic similarity to the original cultivar. The relative expression levels of six genes (i.e., SoDREB, SoNHX1, SoSOS1, SoHKT, SoBADH, and SoMIPS) were found to be significantly higher in the salt-tolerance clones than those measured in the original plant. The measured proline levels, the glycine betaine content, the relative water content, the SPAD unit, the contents of chlorophyll a and b, as well as the K⁺/Na⁺ ratios of the salt-tolerant clones were also found to be significantly higher than those of the original plant.When the salt-tolerant clones were grown in a low saline soil, they exhibited a higher Brix percentage than that of the original cultivar.     

一年生盐生植物耐盐机制研究进展

盐生植物是一类能够在盐土上完成生活史的天然植物, 在与盐土协同演化过程中形成了一系列适应盐生环境的特殊生存策略。其中一年生盐生植物因其生活史短、方便培养和观察、易于基因转化和后代繁殖, 已成为耐盐机制研究的主要对象。一年生盐生植物面临多变的生境胁迫, 具有更大的生存风险, 所以具有不同于多年生盐生植物的更稳妥的适应机制, 主要体现在种子的高盐休眠、复水速萌、形态和萌发的多态性、存在持久种子库及调节资源分配等方面。种子萌发后的生长、发育和繁殖等生活史的各阶段都要经受严峻的盐生胁迫环境。通常所说的耐盐机理是指成株对盐分的调控, 按照植物种类不同而分为稀盐、泌盐和拒盐3种耐盐形式。该文在对国内外相关文献进行分析归纳的基础上, 首先介绍了一年生盐生植物的常见类型, 然后分别从种子特征、形态结构、生理生化和生态习性等方面综述了一年生盐生植物的耐盐机制。     

宏基因组学应用于耐盐酶类及耐盐基因研究的进展

耐盐酶在高盐浓度下仍具备催化活性和稳定性,在高盐食品和海产品加工、洗涤及其它高盐环境生物技术领域被广泛应用;耐盐基因在高盐条件下可以使微生物维持正常功能,获取并研究不同环境中的耐盐基因对揭示微生物的耐盐机制,以及实现其在高盐环境中的定向应用具有的重要意义。宏基因组学避开纯培养技术探知微生物的多样性及其功能,为我们提供了一种发现新基因、开发新的微生物活性物质和研究微生物群落结构及其功能的新技术。文中结合本课题组的研究工作,综述了利用宏基因组学获取耐盐酶类及耐盐基因的策略,同时着重介绍利用宏基因组学从海洋、土壤、胃肠道等环境中获取耐盐酶类及耐盐基因的研究。     

二色补血草的耐盐性研究

通过NaCl胁迫发芽试验、盆栽浇灌NaCl盐水试验、不同含盐量土壤的种植试验,研究二色补血草（Limoniumbicolor）3个时期的耐盐能力。结果表明：高浓度NaCl对二色补血草种子的萌发有明显的抑制作用,种子的发芽率与盐浓度之间呈显著的负相关,种子萌发时盐胁迫的适宜值、临界值、极限值分别是0.49%、1.30%、2.11%;盆栽试验结果表明,二色补血草在2.0%以下的NaCl盐水胁迫下没有表现明显的盐害症状,只表现高浓度NaCl胁迫下,生长量低于低浓度处理的,当处理浓度达到2.4%时,多数植株濒临死亡;不同含盐量土壤的种植试验结果表明,在自然条件下,二色补血草成株耐盐临界值为1.7%～1.8%。试验结果说明二色补血草具有较强的耐盐能力,适合在滩涂盐碱地推广种植。     

二色补血草的耐盐性研究

通过NaCl胁迫发芽试验、盆栽浇灌NaCl盐水试验、不同含盐量土壤的种植试验,研究二色补血草(Limonium bicolor)3个时期的耐盐能力。结果表明:高浓度NaCl对二色补血草种子的萌发有明显的抑制作用,种子的发芽率与盐浓度之间呈显著的负相关,种子萌发时盐胁迫的适宜值、临界值、极限值分别是0.49%、1.30%、2.11%;盆栽试验结果表明,二色补血草在2.0%以下的NaCl盐水胁迫下没有表现明显的盐害症状,只表现高浓度NaCl胁迫下,生长量低于低浓度处理的,当处理浓度达到2.4%时,多数植株濒临死亡;不同含盐量土壤的种植试验结果表明,在自然条件下,二色补血草成株耐盐临界值为1.7%~1.8%。试验结果说明二色补血草具有较强的耐盐能力,适合在滩涂盐碱地推广种植。     

高粱材料耐盐性综合评价方法的初步建立与验证

用0、0.2%、0.4%、0.6%、0.8%5个浓度的NaCl溶液对10份高粱材料进行处理,测定了发芽盐害率、芽高盐害率、侧根数盐害率、根长盐害率、1/2叶片萎蔫持续时间和整个叶片萎蔫持续时间6项指标。以发芽期和苗期的各项指标为基础,根据每个指标盐害率制定各指标的得分方法,最后通过各个材料的得分判断其耐盐性,建立了一套高粱材料的耐盐评价标准。以此标准对10份高粱材料的耐盐性进行评价,排列出这10份高粱材料的耐盐次序,将高粱材料划分为4个耐盐等级。从这10份高粱材料中随机挑选5份耐盐性不同的材料通过盆栽试验对其耐盐性进行鉴定,以检验该评价方法的可靠性,结果表明:盆栽试验的鉴定结果与按照耐盐评分标准得出的结果一致,由此确定此评价方法运用于筛选高粱材料的耐盐性是准确、可靠的。同时在这10份高粱材料中筛选出了耐盐性较强的高粱材料河农1号和能饲1号。     

Molecular cloning and expression analysis of RrNHX1 and RrVHA-c genes related to salt tolerance in wild Rosa rugosa

Salt stress is one important factor influencing the growth and development of plants, and salt tolerance of plants is a result of combined action of multiple genes and mechanisms. Rosa rugosa is not only an important ornamental plant, but also the natural aromatic plant of high value. Wild R. rugosa which is naturally distributed on the coast and islands of China has a good salt tolerance due to the special living environment. Here, the vacuolar Na⁺/H⁺ reverse transporter gene (NHX1) and the vacuolar H⁺-ATPase subunit C gene (VHA-c) closely related to plant salt tolerance were isolated from wild R. rugosa, and the expression patterns in R. rugosa leaves of the two genes under NaCl stress were determined by real-time quantitative fluorescence PCR. The results showed that the RrNHX1 protein is a constitutive Na⁺/H⁺ reverse transporter, the expression of the RrNHX1 gene first increased and then decreased with the increasing salt concentration, and had a time-controlled effect. The RrVHA-c gene is suggestive of the housekeeping feature, its expression pattern showed a similar variation trend with the RrNHX1 gene under the stress of different concentrations of NaCl, and its temporal expression level under 200 mM NaCl stress presented bimodal change. These findings indicated that RrNHX1 and RrVHA-c genes are closely associated with the salt tolerance trait of wild R. rugosa.     

Developing salt tolerant plants in a new century: a molecular biology approach

Soil salinity is a major abiotic stress in plant agriculture strongly, influencing plant productivity world-wide. Classical breeding for salt tolerance in crop plants has been attempted to improve field performance without success. Therefore, an alternative strategy is to generate salt tolerant plants through genetic engineering. Several species and experimental approaches have been used in order to identify those genes that are important for salt tolerance. Due to high level of salt tolerance, halophytes are good candidates to identify salt tolerance genes. However, other species such as yeast and glycophytes have also been employed. Three approaches are commonly used to identify genes important for salt tolerance. The first approach is to identify genes involved in processes known to be critical for salt tolerance (osmolyte synthesis, ion homeostasis, etc.). The second approach is to identify genes whose expression is regulated by salt stress. This is relatively simply and applicable to any plant species. Genetic amenability of some species allows the third approach, which consists in the identification of salt tolerance determinants based on functionality. At the moment, there is a large number of reports in the literature claiming that plants with increased salt tolerance have been obtained. The main problem is that different plant species, stage of development, organs, promoters and salt conditions used it is difficult to compare the degree of salt tolerance conferred by different genes. In this review, we discuss progress made towards understanding the molecular elements involved in salt stress responses that have been used in transgenic approaches to improve salt tolerance.     

Plant abiotic stress: deciphering remedial strategies for emerging problem

Growing in their natural environment, plants often encounter unfavorable environmental conditions that interrupt normal plant growth and productivity. Drought, high/low temperature and saline soils are the most common abiotic stresses that plants encounter in their natural environments. Molecular and genomic analyses have facilitated gene discovery and enabled genetic engineering using several functional or regulatory genes that are known to be involved in stress response and preliminary tolerance, to activate specific or broad pathways related to abiotic stress tolerance in plants. Through the use of transgenic technology, goals such as production of plants with desired traits that were unattainable with traditional selection programs are achieved. This review deals with recent advancement in understanding the role of various stress responsive genes and their critical importance for explaining the control mechanism of abiotic stress tolerance and engineering stress tolerant crops based on the expression of specific stress related genes.     

CBF:平衡植物低温应答与生长发育的关键

低温是影响植物生长发育以及植被分布的重要环境因子。目前,低温信号研究中比较清楚的是CBF依赖的低温信号途径。该文总结了近年来有关CBF的研究成果,详细介绍了CBF家族成员在植物耐寒性中的重要作用,着重分析与讨论CBF介导的低温调控网络及一系列复杂调控机制。理解CBF的复杂作用机制有助于了解植物中CBF介导的冷信号如何平衡耐寒性与生长发育,进而有助于耐寒作物的培育。     

过表达TaLEA1和TaLEA2基因提高转基因拟南芥的耐盐性

我国土壤盐碱化日益严重,对我国的粮食安全造成了严重威胁。耐盐基因挖掘对作物耐盐育种非常重要。LEA蛋白家族是一个多基因家族,在植物应对非生物胁迫中发挥重要作用。本课题组前期研究阐明小麦TaLEA1基因在拟南芥中过表达可以提高转基因植物的耐盐性和抗旱性。本研究系统分析了小麦TaLEA2基因表达蛋白的理化性质、基因表达模式及启动子功能区域,并在拟南芥中过表达TaLEA2基因及共表达TaLEA1和TaLEA2基因,分析TaLEA2基因的抗逆功能及2个LEA基因的抗逆效果。结果表明,TaLEA2基因的表达产物属于第3组LEA蛋白,是稳定的亲水蛋白,富含α-螺旋、β-转角等结构。TaLEA2基因在小麦根、茎、叶、花、种子等不同组织中均有表达,盐胁迫条件诱导其高表达。在拟南芥中过表达TaLEA2基因,或过表达TaLEA1和TaLEA2基因都能够提高转基因拟南芥的耐盐性和抗旱性,转基因株系的种子萌发率、根长及叶绿素含量显著高于野生型,且双基因过表达的转基因植物的抗逆能力高于单个基因过表达株系。本研究结果为LEA基因抗逆机理的研究和多基因共转提高植物抗逆性提供了重要信息。     

小麦TaLEA1基因的表达特征及抗逆功能验证

我国土壤盐碱化日益严重,对我国的粮食安全造成了严重威胁,因此耐盐基因挖掘对作物耐盐育种非常重要。许多研究表明胚胎发育晚期丰富蛋白(LEA)在植物应对非生物胁迫中发挥积极作用。本研究以小麦TaLEA1基因为研究对象,分析了其表达蛋白的理化性质及基因表达模式,并通过在拟南芥中过表达,分析Ta LEA1基因的抗逆功能。结果表明,TaLEA1基因的表达蛋白属于第3组LEA蛋白,是稳定的亲水蛋白,富含α-螺旋、β-转角等结构。Ta LEA1基因在小麦根、茎、叶、花、种子等不同组织中均有表达,盐胁迫条件诱导其高表达。在拟南芥中过表达TaLEA1基因,显著提高了盐胁迫下转基因拟南芥的种子萌发率、根长及盐和旱胁迫下的叶绿素含量。本研究结果为LEA基因抗逆机理的研究和耐盐基因的挖掘提供了重要信息。     

耐盐转基因植物研究进展

高盐是限制作物生长、发育和产量的最严重的非生物胁迫之一。长期以来,改善作物的耐盐性一直是一个伟大的目标。然而,由于耐盐反应是一个极为复杂的过程,过去,通过传统的育种和遗传工程取得的成功有限。近十年来,由于分子生物学的发展,发现了一些与耐盐相关的新基因,对于这些基因的表达方式及其在耐盐反应中的作用已逐步得到了解,这为转基因工程提供了新的材料。通过控制耐盐相关基因在植物体内的表达,已获得了一些提高耐盐性的转基因植物,展示了诱人的前景,但该领域研究仍然存在许多困难和问题,文章重点讨论耐盐转基因植物的进展。