水稻耐盐基因定位与克隆及品种耐盐性分子标记辅助选择改良研究进展

土壤盐渍化严重制约水稻生产发展,提高耐盐性已成为水稻育种的重要目标之一。挖掘水稻耐盐新基因,解析其分子作用机制可以为水稻耐盐性遗传改良奠定基础。本文从定位群体、耐盐性鉴定时期和鉴定方法、耐盐性评价指标、鉴定到的耐盐QTL、耐盐QTL的精细定位和图位克隆等方面,总结了近年来水稻耐盐QTL定位研究中所取得的进展;介绍了水稻耐盐/盐敏感突变体筛选和基因克隆以及耐盐性关联分析的研究近况;并对水稻耐盐性分子标记辅助选择改良的现状作了概述。     

水稻耐盐育种研究进展

盐胁迫对作物的不利影响使作物生长受到抑制,进而影响产量和品质.水稻也是一样,其产量和品质也会受到盐分胁迫的不利影响.从耐盐水稻品种的选育、水稻耐盐的分子机制、耐盐基因的克隆、现代生物技术与耐盐水稻育种方面进行阐述,以期有助于表征未知的耐盐性机制并促进育种应用.     

盐胁迫对水稻的影响及水稻耐盐育种研究

盐胁迫是盐渍化地区水稻生产的主要制约因素之一。盐胁迫对水稻的形态发育及生理生化代谢产生不利影响,其作用机理主要表现在渗透胁迫和离子毒害等。耐盐水稻品种的选育方法主要是筛选耐盐材料,将常规选育和转基因、分子标记辅助选择等现代生物技术相结合,选育耐盐性强且具有实用价值的品种。     

粳稻种质资源芽期耐盐性综合评价与筛选

【目的】土壤盐渍化是危害水稻生产的重要非生物胁迫之一。鉴定水稻种质资源发芽期耐盐性,筛选耐盐指标,培育耐盐品种,对水稻生产的发展具有重要意义。【方法】利用125 mmol/L Na Cl溶液对64份粳稻种质资源进行盐胁迫,于胁迫后3 d测定发芽数;胁迫5 d、10 d后,测定发芽数、芽长和根长,并计算相对芽长、根长、发芽势、发芽率、盐害率,发芽指数和活力指数。运用多种统计学方法对各种质资源的芽期耐盐性进行综合评价,分析典型耐盐和盐敏感种质盐胁迫条件下的发芽特征。【结果】相对盐害率与相对根长、相对发芽势、相对发芽率、发芽指数和活力指数均极显著负相关;除相对芽长外各指标间的相关性均达到极显著水平。通过聚类分析将64份粳稻种质资源划分成4个类群。第Ⅰ、Ⅰ类群分别为典型的盐敏感和耐盐类群,第Ⅱ类群为弱耐盐种质为主的混合类群,第Ⅲ类群主要由耐盐种质组成。通过主成分分析将7个评价指标转换为3个主成分,应用隶属函数和权重,获得了客观评价粳稻种质资源耐盐性的综合评价值D。分别选取D值最高和最低的5份种质资源进行芽期耐盐指标的差异显著性分析,结果表明,两组种质资源盐胁迫5 d的各评价指标差异均达到极显著水平,10 d的评价指标除相对芽长外,均达到了显著差异水平。【结论】水稻芽期对盐胁迫较为敏感,且耐盐性不同的种质间差异显著。利用逐步回归和主成分分析获得发芽指数、相对根长和相对盐害率3个指标,可作为快速鉴定粳稻种质资源芽期耐盐性的重要指标,若采用多元统计方法评价可靠性更高。     

水稻耐盐育种研究进展

培育耐盐水稻品种是盐碱地的粮食作物增产和对盐碱地改良的重要途径之一。水稻对盐胁迫中度敏感,其耐(敏)盐性状是受多基因控制的数量性状,易受环境条件等因素影响。到2015年,利用AFLP、RFLP、SSR等分子标记手段在水稻12条染色体上定位的耐盐QTL已有250多个,其中以第1、2、6和7染色体上居多,但克隆的水稻耐盐基因相对较少。水稻耐盐品种的选育现仍以常规育种为主,即主要通过筛选耐盐种质及耐盐鉴定,再利用杂交和回交等方法将耐盐基因导入到优良水稻品种中,从而选育出综合性状优良的耐盐品种。本文阐述了国内外水稻种质耐盐性鉴定技术和指标、耐盐种质筛选、耐盐品种选育以及耐盐性的遗传及分子机理等研究进展,并对今后水稻耐盐育种研究工作提出了展望与建议。     

NaCl胁迫对不同耐盐性水稻某些生理特性和光合特性的影响

选取较耐盐的水稻品种‘HH11’、‘JX99’和盐敏感水稻品种‘YSXD’,设置6个土壤NaCl浓度处理(0、1、2、3、4 g/kg),在防雨棚下盆栽并培育至孕穗期,分析NaCl胁迫对不同耐盐性水稻生理生化及光合特性的影响,结果表明:(1)NaCl胁迫抑制水稻的生长,表现为随着NaCl浓度增加,水稻的株高逐渐降低,但在3～4 g/kg土壤含盐量下耐盐水稻的株高显著高于盐敏感水稻品种。(2)耐盐水稻和盐敏感水稻的可溶性总糖对NaCl胁迫的响应差异明显,在1～4 g/kg NaCl胁迫下,盐敏感水稻叶片可溶性总糖显著降低,但是耐盐水稻可溶性总糖大量合成并积累,并且显著高于盐敏感品种。(3)NaCl浓度增加迫使水稻叶片丙二醛不断积累,导致细胞膜透性逐渐增大,但是耐盐水稻丙二醛的积累量较少,细胞膜受盐害程度显著小于盐敏感水稻。(4)NaCl胁迫抑制了水稻光合速率,但不同的NaCl浓度下导致水稻品种光合速率下降的原因各有差异,其中在0～1 g/kg NaCl胁迫下盐敏感水稻光合速率降低是非气孔因素导致的,而在2～4 g/kg NaCl胁迫下是由气孔因素造成的;0～2g/kg盐浓度下耐盐水稻HH11的光合速率降低是非气孔因素导致的,3～4 g/kg盐浓度处理是由气孔因素导致的;气孔因素是0～4 g/kg盐处理耐盐水稻JX99的光合速率降低的主要原因。(5)盐胁迫下耐盐水稻的叶片蒸腾速率显著降低,并且显著低于盐敏感水稻,相反水分利用效率和气孔限制值却明显升高,并且显著高于盐敏感水稻品种,表明盐浓度增加迫使耐盐水稻气孔阻力增大,减少水分的流失,抑制了蒸腾速率,使耐盐水稻叶片保持较高的水势;同时提高了叶片水分利用效率,碳同化效率提高,以满足耐盐水稻正常代谢生理需求,进行正常生命活动。     

棉花耐盐性研究现状与展望

从棉花耐盐特性、盐胁迫对棉花植株生长发育的影响、耐盐机理和棉花耐盐性的提高与调控四个方面进行了综述,并对棉花耐盐性研究的发展方向进行了展望。     

耐盐性不同水稻品种对盐胁迫的响应及其生理机制

【目的】阐明耐盐性不同水稻品种对盐胁迫的响应差异及其生理机制。【方法】耐盐水稻品种连鉴5号、盐稻16Z38和盐敏感水稻品种华粳5号、连粳17号生长于盆钵,盆栽土设置2个不同盐浓度处理：0 g/kg (对照);3 g/kg(盐胁迫)。【结果】与对照相比,在盐胁迫下两类品种均有不同程度的减产,耐盐品种减产的幅度明显小于盐敏感品种。耐盐水稻品种具有较高的产量主要在于较高的颖花量和结实率。与盐敏感品种相比,耐盐水稻品种在分蘖中期、拔节期、抽穗期和灌浆中期叶片中超氧化物歧化酶等抗氧化酶活性较高,拔节期和抽穗期具有较高根系脯氨酸含量、脯氨酸合成酶活性和较高的K⁺/Na⁺值,分蘖至拔节以及抽穗至成熟期具有较高的作物生长率,抽穗期具有较高的叶面积指数和叶片SPAD值,抽穗至灌浆中期具有较高的根系氧化力。【结论】在盐胁迫下耐盐水稻品种较高的总颖花量、较强的抗氧化和渗透调节能力、较高的根系氧化力和K⁺/Na⁺值是其获得较高产量的重要原因。在盐胁迫下叶片超氧化物歧化酶活性较强和根系氧化力较高可作为评定水稻品种耐盐性的重要...     

过表达一个疣粒野生稻的Ricin_B凝集素基因OmR40c1增强水稻耐盐性

盐胁迫是影响盐/碱稻区水稻产量的主要逆境因素。研究耐盐相关基因,对于培育水稻抗逆品种具有重要意义。OmR40c1是从疣粒野生稻(Oryza meyeriana)中分离获得cDNA序列,与已报道的受脱落酸与盐诱导的OsR40c1的CDS序列完全相同,但在5′端UTR和3′端UTR区域有差别。有关OmR40c1基因功能的研究还未见报道。本研究的主要目的是初步分析OmR40c1在水稻耐盐性中的作用。对栽培稻日本晴(Oryza sativa L.var.Nipponbare)进行脱落酸(abscisic acid,ABA)和盐胁迫处理,OsR40c1基因均上调表达。构建基因定位载体并注射烟草表皮细胞,其结果表明OmR40c1定位于细胞质膜和细胞核上。构建超表达载体并转染日本晴,PCR及qRT-PCR结果均显示OmR40c1在转基因材料中成功表达。OmR40c1可能参与种子萌发。174mmol/L盐胁迫下,野生型种子的发芽率下调一半左右,转OmR40c1基因水稻种子发芽未受影响;转OmR40c1水稻苗期的株高是未经盐处理转基因水稻株高的一半左右;而野生型的株高是未经盐处理野生型水稻株高的1/6左右。盐胁迫下,水稻叶片、根长等都受到不同程度的影响。盐胁迫下,转OmR40c1基因水稻植株内的Na~+浓度是野生型的1.38倍;K~+浓度是野生型的1.25倍。综上,OmR40c1一定程度提高了水稻的耐盐性,且苗期的耐盐性高于成株期。     

盐胁迫下 4 个不同耐盐基因型水稻 Na+ 、K+ 积累效应

选取耐盐性较强的水稻株系‘Fl478’、‘JX99’、‘Pokkali’和盐敏感型品种‘IR29’,设置 6 个 NaCl 浓度梯度处理, 分别为 0、0.1%、0.2%、0.3%、0.4%、0.5%,采用桶栽土培的方法培育至孕穗期,研究盐胁迫对孕穗期水稻植株不同 器官 Na+ 、K+ 分配规律与积累的影响。结果表明：（1）盐胁迫下水稻株系不同器官 Na+ ,K+ 的积累效应存在差异,水稻 株系耐盐性的差异主要表现在根系,而叶鞘对 Na+ 、K+ 的吸收与分配的差异明显。（2）耐盐植株根系聚 Na+ 能力较强, 叶片、叶鞘积累较多的 K+ ,而感盐品种根系积累 K+ 和 Na+ 以缓解根系渗透胁迫,调节根系 Na+ /K+ 平衡稳态,保持水稻 正常的代谢活动。（3）叶鞘是水稻植株关键的 Na+ -K+ 调库,通过吸收和分配 Na+ 来调节根、叶、叶鞘 Na+ /K+ 平衡以提 高水稻耐盐性。（4）耐盐株系叶鞘向叶片选择运输 K+ 的能力、限 Na+ 运输能力强于盐敏感品种,叶鞘对 Na+ 、K+ 吸收 与分配运输能力的大小,决定水稻株系的耐盐性,具体表现耐盐品种叶鞘向吸收与运输 K+ 能力较强、根系限 Na+ 能力 显著高于感盐植株。（5）0.4%~0.5%盐浓度限制水稻叶鞘 Na+ -K+ 库吸收与分配能力,各组织中 Na+ /K+ 失衡,植株耐盐 性降低,受盐害程度加深。（6）盐胁迫促进水稻株系叶鞘向根系吸收较多 Na+ 和向叶片、根系输出 K+ ,却限制 Na+ 对 叶片的分配,保证根系保持较高的 K+ /Na+ ,耐盐幼嫩叶片积累较多的 K+ 以维持叶片组织保卫细胞渗透平衡,保证水稻 植株获得生活必需的光合原料,而盐敏感植株积累的 K+ 主要用于缓解组织渗透胁迫,以维持正常生命活动。     

Antioxidant Defense Mechanisms of Salinity Tolerance in Rice Genotypes

In order to elucidate the role of antioxidant responses in salinity tolerance in rice genotypes under salt stress, experiments were conducted using four rice varieties, including salt-sensitive BRRI dhan 28 and three salt-tolerant varieties BRRI dhan 47, BINA dhan 8 and BINA dhan 10. Thirty-day-old rice seedlings were transplanted into pots. At the active tillering stage(35 d after transplanting), plants were exposed to different salinity levels(0, 20, 40 and 60 mmol/L NaCl). Salt stress caused a significant reduction in growth for all the rice genotypes. Growth reduction was higher in the salt-sensitive genotype than in the salt-tolerant ones, and BINA dhan 10 showed higher salt tolerance in all measured physiological parameters. The reduction in shoot and root biomass was found to be minimal in BINA dhan 10. Chlorophyll content significantly decreased under salt stress except for BINA dhan 10. Proline content significantly increased in salt-tolerant rice genotypes with increased salt concentration, and the highest proline content was obtained from BINA dhan 10 under salt stress. Catalase and ascorbate peroxidase activities significantly decreased in salt-sensitive genotype whereas significantly increased in salt-tolerant ones with increasing salt concentration. However, salt stress significantly decreased guaiacol peroxidase activity in all the rice genotypes irrespective of salt tolerance. K~+/Na~+ ratio also significantly decreased in shoots and roots of all the rice genotypes. The salt-tolerant genotype BINA dhan 10 maintained higher levels of chlorophyll and proline contents as well as catalase and ascorbate peroxidase activities under salt stress, thus, this might be the underlying mechanism for salt tolerance.     

盐胁迫对粳稻品种生长和生理特性的影响

以津原85、金穗26、隆粳27、辽粳763为试验材料,研究了不同盐胁迫浓度对这4个品种生长和抗逆性的影响,以期为盐碱地适宜水稻品种的筛选提供理论参考.结果 表明,盐胁迫显著抑制水稻幼苗的生长,金穗26受到的影响最小.在盐胁迫下,津原85离子选择吸收性较强,有利于提高耐盐性.盐胁迫对水稻的主要伤害是破坏膜质结构,进而影响...     

Identification of Rice Accessions Associated with K~+/Na~+ Ratio and Salt Tolerance Based on Physiological and Molecular Responses

The key for rice plant survival under Na Cl salt stress is maintaining a high K~+/Na~+ ratio in its cells. Selection for salt tolerance rice genotypes based on phenotypic performance alone will delay in progress in breeding. Use of molecular markers in tandem with physiological studies will help in better identification of salt tolerant rice accessions. Eight rice accessions along with the check Dongjin were screened using 1/2 Yoshida solution with 50 mmol/L NaCl at the seedling stage. The accessions IT001158, IT246674, IT260533 and IT291341 were classified as salt tolerant based on their K~+/Na~+ ratios. Seventeen SSR markers reported to be associated with K~+/Na~+ ratio were used to screen the accessions. Five SSR markers(RM8053, RM345, RM318, RM253 and RM7075) could differentiate accessions classified based on their K~+/Na~+ ratios. Banding pattern of the accessions was scored compared to the banding pattern of Dongjin. The study differentiated accessions based on their association of K~+/Na~+ ratio with molecular markers which are very reliable. These markers can play a significant role in screening large set of rice germplasms for salt tolerance and also help in identification of high-yielding varieties with better salt tolerance. The salt tolerant accessions can be taken forward into developing better varieties by conventional breeding and exploring genes for salt tolerance.     

盐胁迫对水稻幼苗期的影响研究——盐胁迫对水稻干物质的影响

对123个不同水稻品种分别在纯水、100 mmol/L NaCl和150 mmol/LNaCl水溶液中的萌发情况进行试验,来研究盐对幼苗单株苗干重,去苗后的种子干重,种子的贮藏物质利用率的影响规律,及利用率对盐胁迫的不同反应,结果表明:在盐胁迫下不同品种单株苗干重显著下降。盐浓度增高,单株苗干重下降率增大。去苗后的种子干重在盐胁迫下明显的升高,盐浓度增大,去苗后的种子干重也增大。在盐胁迫下,各品种的种子贮藏物质利用率都下降。盐浓度增大,种子贮藏物质利用率降低。     

Inheritance and QTL Mapping of Salt Tolerance in Rice

An F2 population derived from the cross between Jiucaiqing (japonica) and IR36 (indica) was used to analyze the inheritance of salt tolerance in rice by genetic model of major-genes plus polygenes, and to map the corresponding QTLs by SSR molecular markers. Rice plants of P1, P2, F1 and F2 at 5- to 6- leaf stage were treated under 140 mmol/L NaCI for 10 days. Three indices representing the ability of salt tolerance of rice seedlings were measured, including salt tolerance rating (STR), Na^ /K^ ratio in roots and dry matter weight of shoots (DWS). STR, Na^ /K^ and DWS were all controlled by two major genes with modification by polygenes. Heritability of these traits from major genes was 17.8, 53.3 and 52.3%, respectively. The linkage map constructed by 62 SSR molecular markers covered a total length of about 1 142 cM. There were three QTLs detected for STR located on chromosome 1, 5 and 9, two QTLs for DWS on chromosomes 8 and 9, and two QTLs for Na^ /K^ on chromosomes 2 and 6, one on each chromosome respectively. Single QTL accounted for 6.7 to 19.3% of phenotypic variation. Identification method of salt tolerance in rice and breeding of rice varieties with salt tolerance based on molecular markers assisted selection had been discussed.     

水稻不同品种耐盐限度研究初报

了解水稻不同品种的耐盐限度,对于盐渍土区稻作生产的持续、稳定发展有着重要意义。试验表明,土壤全盐5.94g/kg时,盐粳68、花粳8、辽粳294和盐粳34的发芽率均不低于90%。花粳15分蘖期最高耐盐为14g/kg,盐粳68拔节孕穗期最高耐盐为17g/kg,盐丰47-6成熟期最高耐盐为12g/kg。12个供试水稻品种,其中盐粳68、花粳8、辽粳9和92-71耐盐能力强;花粳15、抗盐100、盐丰47-6、辽粳294、盐粳34和盐粳31耐盐能力次之。     

Salt Tolerance in Rice:Focus on Mechanisms and Approaches

Salt tolerance is an important constrain for rice, which is generally categorized as a typical glycophyte. Soil salinity is one of the major constraints affecting rice production worldwide, especially in the coastal areas. Susceptibility or tolerance of rice plants to high salinity is a coordinated action of multiple stress responsive genes, which also interacts with other components of stress signal transduction pathways. Salt tolerant varieties can be produced by marker-assisted selection or genetic engineering by introducing salt-tolerance genes. In this review, we have updated on mechanisms and genes which can help in transferring of the salt tolerance into high-yielding rice varieties. We have focused on the need for integrating phenotyping, genomics, metabolic profiling and phenomics into transgenic and breeding approaches to develop high-yielding as well as salt tolerant rice varieties.     

Genetic diversity and association mapping for salinity tolerance in Bangladeshi rice landraces

Breeding for salinity tolerance using Bangladeshi rice landraces and understand genetic diversity has been limited by the complex and polygenic nature of salt tolerance in rice genotypes. A genetic diversity and association mapping analysis was conducted using 96 germplasm accessions with variable response to salt stress at the seedling stage. These included86 landraces and 10 indica varieties and lines including Nona Bokra, from southern Bangladesh. A total of 220 alleles were detected at 58 Simple Sequence Repeat(SSR) marker loci randomly distributed on all 12 rice chromosomes and 8 Sequence Tagged Site(STS) markers developed for genes SKC1, DST, and SalT. The average gene diversity was 0.5075 and polymorphism information content value was 0.4426, respectively. Cluster analysis revealed that 68 and 21 accessions were clustered into 2 distinct groups, possibly corresponding to indica and japonica groups, respectively and the remaining 7 landraces were classified as an admixed group. In addition to Wn11463, the STS marker for SKC1, RM22418 on Chr. 8 was significantly associated with salinity tolerance, at the location of a QTL detected in previous studies. Our findings of favorable alleles associated with salinity tolerance in Bangladeshi rice landraces, as well as the development of STS markers for salt tolerance genes, will be helpful in future efforts to breed salinity tolerance in rice.     

Stress-Activated Protein Kinase OsSAPK7 Regulates Salt-Stress Tolerance by Modulating Diverse Stress-Defensive Responses in Rice

Soil salinity is an environmental threat limiting rice productivity. Identification of salinity tolerance genes and exploitation of their mechanisms in plants are vital for crop breeding. In this study, the function of stress-activated protein kinase 7(OsSAPK7), a SnRK2 family member, was characterized in response to salt stress in rice. Compared with variety 9804, OsSAPK7-overexpression plants had a greater survival rate, increased chlorophyll and proline contents, and superoxide dismutase and catalase activities at the seedling stage under salt-stress conditions, as well as decreased sodium potassium ratio(Na~+/K~+) and malondialdehyde contents. After salt stress, the OsS APK7 knockout plants had lower survival rates, increased Na~+/K~+ ratios and malomdiadehyde contents, and decreased physiological parameters compared with 9804. These changes in transgenic lines suggested that OsSAPK7 increased the salt tolerance of rice by modulating ion homeostasis, redox reactions and photosynthesis. The results of RNA-Seq indicated that genes involved in redox-dependent signaling pathway, photosynthesis and zeatin synthesis pathways were significantly down-regulated in the OsSAPK7 knockout line compared with 9804 under salt-stress condition, which confirmed that OsSAPK7 positively regulated salt tolerance by modulating diverse stress-defensive responses in rice. These findings provided novel insights for the genetic improvement of rice and for understanding the regulatory mechanisms of salt-stress tolerance.