植物抗寒冻基因工程研究进展

低湿寒害是限制农作物产量和分布的一种全球性的自然灾害。提高农作物的抗寒性具有重要意义。目前随着植物寒害机理、抗寒冻和冷驯化分子机理的深入发展,已研究发现了多种抗寒基因,包括各种抗寒调控基因和各种抗寒功能基因,从而使植物抗寒冻基因工程的研究与应用得以了广泛开展,以期最终有效地提高农作物的抗寒性,增加农业产量。本文综合概述了国内外有关植物抗寒冻基因工程的最新研究方向、进展及成就,并提出了此领域尚存在的一些问题及其前景展望。     

柑桔抗寒研究的现状与展望

柑桔是一种经济价值极高但又常受冻害影响的亚热带果树,本文总结了柑桔抗寒研究取得的成果,介绍了对于柑桔抗寒研究还是空白但是在其他植物抗寒研究上取得的新进展,并由此提出了柑桔抗寒可在冷诱导蛋白、冷诱导基因和抗寒基因工程等方面进行探讨以及作者粗浅的研究设想。     

植物抗寒性研究进展

低温是限制植物生长和分布的一种非生物胁迫因素。因此,提高植物的抗寒性对农业具有十分重要的意义。综述了植物抗寒性的最新进展,包括植物抗寒性机理研究,提高植物抗寒力的方法研究和应用情况;提出了在植物抗寒性研究中存在的问题,并对未来工作重点进行了展望。     

Temperature and cardiovascular deaths in Montreal

The incidence of mortality from myocardial infarction over a 2-year period in Montreal is examined in relation to temperatures and snowfall. Mortality is seen to increase with deviations from seasonally determined thermally neutral conditions, although compared to observations of a parallel study in subtropical Brisbane, death rates are lower with similar falls in temperature. In Montreal, mortality is also seen to increase with snow during the previous day. During anomalous cold spells death frequencies decrease, a phenomenon interpreted as the behavioural thermoregulatory process of cold avoidance.     

LTP在植物抗环境胁迫中的作用

脂质转移蛋(白Lipid transfer protein,LTP)是一类分泌蛋白,曾被认为是一种在体外膜间进行脂质转移的蛋白,因其作用对象很广泛,所以目前又称其为非特异性LTP(non-specific lipid transfer proteins,nsLTPs)。有很多证据表明nsLTPs可能参与多方面的植物细胞与生理生化反应,这些生物学功能包括:参与角质层的合成和胚胎的发育;适应各种胁迫环境;抗病原微生物等作用,尤其是在适应胁迫环境中,起到很大作用。介绍了nsLTPs在植物抗环境胁迫中所发挥的作用,同时对应用研究作了简单展望。     

Effects of a Daily Short-Term Temperature Drop on Chilling-Sensitive and Cold-Resistant Plants

Russian Journal of Plant Physiology - A set of physiological and biochemical parameters (leaf growth rate, plant biomass, chlorophyll content, rates of photosynthesis and transpiration, relative...     

Molecular Basis of Disease Resistance Acquired through Cold Acclimation in Overwintering Plants

Plants require substantial resistance against freezing and pathogens for overwintering. These two traits are acquired through cold acclimation. In contrast to freezing tolerance, molecular basis of disease resistance acquired through cold acclimation is poorly understood. Recent studies have suggested that pathogenesis-related (PR) proteins that are secreted into the apoplast during cold acclimation are responsible for the disease resistance. Interestingly, some of the cold-induced PR proteins display both antifungal and antifreeze activities, suggesting a dual function in protecting plants from overwintering stresses. The signaling pathway for cold-induced disease resistance is currently unknown but can be independent of pathogen-induced defense mechanisms.     

Breeding for Salinity Tolerance in Plants

Accumulation of high levels of salts in the soil is characteristic of arid and semi-arid regions. Although different curative and management measures are being used to render salt-affected soils fit for agriculture, they are extremely expensive and do not provide permanent solutions to overcome the salinity problem. In contrast, a biotic approach for overcoming salinity stress has gained considerable recognition within the past few decades in view of the vast experimental evidence from what has happened in nature concerning the evolution of highly salt-tolerant ecotypes of different plant species, and also from the remarkable achievements that have been made in improveing different agronomic traits through artificial selection.

Considerable improvements in salt tolerance of important crop species have been achieved in the past 2 decades using barley, rice, pearl millet, maize, sorghum, alfalfa, and many grass species. Such achievements relied solely on assessment of the phenotypic expression of the features involved. Knowledge of the underlying physiological mechanisms producing those salt-tolerant individuals was not clearly known. The present review highlights the relationships between different physiological/biochemical variables being recommended as selection criteria, and salt tolerance of different plant species. This paper also lists different sources of genetic variation for salt tolerance since it is evident that for successful improvement in a character there must be a great amount of genetic variation present in the gene pool of a species.     

测定植物抗寒性的电阻抗图谱法

电阻抗图谱(electrical impedance spectroscopy,EIS)分析作为测定植物抗寒性的一种方法,在农业、林业和园艺领域的应用正在不断扩大。该文从EIS的原理入手,讨论了影响电阻抗特性的生理和物理因子;介绍了测定EIS适用的模型;阐述了用EIS测定抗寒性的方法。在EIS分析中,胞外电阻率(re)是确定抗寒性最适用的一个参数,弛豫时间(τ1)是精确度最高的参数。     

柑橘抗寒细胞变异体的获得及其抗性遗传稳定性的研究

锦橙（Citrussinensis（L．）Osbeckcv．Jincneng）珠心愈伤组织悬浮细胞通过γ射线诱变与高浓度羟脯氨酸离体选择的方法,获得了抗性细胞系,并再生植株。抗性细胞系及其再生植株比供体（对照）的抗寒性分别提高1．4℃和24℃,连续3年测定结果表明再生植株抗寒性的增强是稳定的。其叶片总DNA的RAPD分析说明供体与变异植株可能在DNA水平上存在细微的差异,冷驯化进一步加强抗性细胞系与再生植株在抗氧化酶SOD、过氧化氢酶（CAT）活性和脯氨酸（Pro）、丙二醛含量上的差异。变异系抗性增强的生理基础涉及Pro代谢外的其他途径,抗氧化酶活性的提高与其抗寒性的增强有关。     

Salicylhydroxamic Acid-Resistant and Sensitive Components of Respiration in Chilling-Sensitive Plants Subjected to a Daily Short-Term Temperature Drop

Russian Journal of Plant Physiology - Salicylhydroxamic acid (SHAM) is used in studies of plant respiratory metabolism as a specific inhibitor of alternative cyanide-resistant oxidase (AOX). The...     

Enhanced Tolerance to Ascochyta Blight in Chickpea Plants via Low Temperature Acclimation

Russian Journal of Plant Physiology - Low temperature (LT) and Ascochyta blight are two major stresses in chickpea (Cicer arietinum L.) cultivation. After exposure to LT treatments (acclimation,...     

Cold resistance mechanisms in high desert Andean plants

Freezing tolerance and freezing avoidance were studied, during the growing season, in plant species from two different elevations (3200 m and 3700 m) in a desert region of the high Andes (29° 45S, 69° 59W) in order to determine whether there was a relationship between plant height and cold resistance mechanisms. Freezing injury and supercooling capacity were determined in plants of different height, from ground-level (<20 cm tall) to tall shrubs (27–90 cm). All ground-level plants showed freezing tolerance as the main mechanism for resistance to freezing temperatures. Tall shrubs avoided freezing temperatures, mainly through supercooling. Supercooling was only present in plants occupying the lower elevation (i.e., 3200 m). Both avoidance and tolerance mechanisms are present in a single genus (i.e., Adesmia).     

Impact of Water Availability on Responses of Cucumis sativus Plants to a Short-Term Daily Temperature Drop

Russian Journal of Plant Physiology - Impact of relative air humidity (RAH) and watering mode on responses of cucumber (Cucumis sativus L.) plants to a daily short-term (2 h) temperature decrease...     

Chlorophyll Fluorescence as a Possible Tool for Salinity Tolerance Screening in Barley (Hordeum vulgare L.)

The application of chlorophyll fluorescence measurements to screening barley (Hordeum vulgare L.) genotypes for salinity tolerance has been investigated. Excised barley leaves were cut under water and incubated with the cut end immersed in water or in a 100-mM NaCl solution, either in the dark or in high light. Changes in rapid fluorescence kinetics occurred in excised barley leaves exposed to the saline solution only when the incubation was carried out in the presence of high light. Fluorescence changes consisted of decreases in the variable to maximum fluorescence ratio and in increases in the relative proportion of variable fluorescence leading to point I in the Kautsky fluorescence induction curve. These relative increases in fluorescence at point I appeared to arise from a delayed plastoquinone reoxidation in the dark, since they disappeared after short, far-red illumination, which is known to excite photosystem I preferentially. We show that a significant correlation existed between some fluorescence parameters, measured after a combined salt and high-light treatment, and other independent measurements of salinity tolerance. These results suggest that chlorophyll fluorescence, and especially the relative fluorescence at point I in the Kautsky fluorescence induction curve, could be used for the screening of barley genotypes for salinity tolerance.     

A Calcium/Calmodulin-regulated Member of the Receptor-like Kinase Family Confers Cold Tolerance in Plants

Cold is a limiting environmental factor that adversely affects plant growth and productivity. Calcium/calmodulin-mediated signaling is believed to play a pivotal role in plant response to cold stress, but its exact role is not clearly understood. Here, we report that CRLK1, a novel calcium/calmodulin-regulated receptor-like kinase, is crucial for cold tolerance in plants. CRLK1 has two calmodulin-binding sites with different affinities as follows: one located at residues 369–390 with a K_d of 25 nm, and the other located at residues 28–112 with a K_d of 160 nm. Calcium/calmodulin stimulated the kinase activity, but the addition of chlorpromazine, a calmodulin antagonist, blocked its stimulation. CRLK1 is mainly localized in the plasma membrane, and its expression is stimulated by cold and hydrogen peroxide treatments. Under normal growth conditions, there is no noticeable phenotypic difference between wild-type and crlk1 knock-out mutant plants. However, as compared with wild-type plants, the crlk1 knock-out mutants exhibited an increased sensitivity to chilling and freezing temperatures. Northern analysis showed that the induction of cold-responsive genes, including CBF1, RD29A, COR15a, and KIN1 in crlk1 mutants, is delayed as compared with wild-type plants. These results indicate that CRLK1 is a positive regulator of cold tolerance in plants. Furthermore, our results suggest that CRLK1 plays a role in bridging calcium/calmodulin signaling and cold signaling.     

中国水稻微核心种质不同生育时期耐冷性鉴定及其相关分析

以204份中国水稻微核心种质为试验材料,进行了水稻发芽期、芽期、幼苗期、孕穗期等耐冷性鉴定及其相关性分析。结果表明,水稻各生育时期耐冷性在籼粳亚种间、各种质之间存在明显的差异。粳稻种质的耐冷性明显强于籼稻种质,但籼稻种质中也存在耐冷性较强的种质。高阳淀稻大红芒、肥东塘稻、木樨球、卫国、兴国、山酒谷、中花8号等粳稻种质和包选21号、红米三担白、寸谷糯、红谷等籼稻种质在水稻各生育时期均表现较强的耐冷性,在水稻耐冷性育种及耐冷基因发掘研究中应加以利用。自然低温和冷水胁迫下水稻结实率与低温下发芽率呈显著或极显著正相关,而与死苗率呈显著负相关,即低温下发芽期耐冷性和芽期耐冷性强的水稻种质一般表现为较强的孕穗期耐冷性。认为低温下发芽率和芽期低温处理后的死苗率可以作为孕穗期耐冷性早期鉴定的间接指标。     

A Subset of Cytokinin Two-component Signaling System Plays a Role in Cold Temperature Stress Response in Arabidopsis

A multistep two-component signaling system is established as a key element of cytokinin signaling in Arabidopsis. Here, we provide evidence for a function of the two-component signaling system in cold stress response in Arabidopsis. Cold significantly induced the expression of a subset of A-type ARR genes and of GUS in Pro_ARR7:GUS transgenic Arabidopsis. AHK2 and AHK3 were found to be primarily involved in mediating cold to express A-type ARRs despite cytokinin deficiency. Cold neither significantly induced AHK2 and AHK3 expression nor altered the cytokinin contents of wild type within the 4 h during which the A-type ARR genes exhibited peak expression in response to cold, indicating that cold might induce ARR expression via the AHK2 and AHK3 proteins without alterations in cytokinin levels. The ahk2 ahk3 and ahk3 ahk4 mutants exhibited enhanced freezing tolerance compared with wild type. These ahk double mutants acclimated as efficiently to cold as did wild type. The overexpression of the cold-inducible ARR7 in Arabidopsis resulted in a hypersensitivity response to freezing temperatures under cold-acclimated conditions. The expression of C-repeat/dehydration-responsive element target genes was not affected by ARR7 overexpression as well as in ahk double mutants. By contrast, the arr7 mutants showed increased freezing tolerance. The ahk2 ahk3 and arr7 mutants showed hypersensitive response to abscisic acid (ABA) for germination, whereas ARR7 overexpression lines exhibited insensitive response to ABA. These results suggest that AHK2 and AHK3 and the cold-inducible A-type ARRs play a negative regulatory role in cold stress signaling via inhibition of ABA response, occurring independently of the cold acclimation pathway.     

野生黄花苜蓿冷诱导基因克隆及序列分析

根据紫花苜蓿抗寒基因cas15B(登录号:L12462)的cDNA序列,设计1对特异引物,以经过低温胁迫的黄花苜蓿总RNA为模板,采用RT-PCR方法克隆获得了550bp的cDNA片段。测序和序列分析结果表明,扩增片段长度为550bp,编码159个氨基酸。主要由Gly(甘氨酸)、Glu(谷氨酸)、His(组氨酸)、Lys(赖氨酸)这4种氨基酸组成,占总量的70%。含1个重复了5次的10肽基序,其序列为Lys-Gly-Glu-Gln-His-Gly-His(Phe)-Val(Leu)-Gly-Gly。经序列比较分析,该片段与紫花苜蓿冷诱导基因CAS15B的核苷酸、氨基酸的同源性均为90%,命名为MfCAS15-1。亚细胞结构定位分析结果显示,MfCAS15-1是一种定向到核的蛋白,在调节或维持核的结构与功能方面起作用。本研究在黄花苜蓿中成功获得了抗寒基因同源序列,为最终克隆黄花苜蓿MfCAS15-1抗寒基因全长奠定了基础。