Seedling and adult plant resistance to leaf rust in 46 Chinese bread wheat landraces and 39 wheat lines with known Lr genes

Wheat leaf rust,caused by Puccinia triticina(Pt),is an important foliar disease that has an important influence on wheat yield.The most economic,safe and effective way to control the disease is growing resistant cultivars.In the present study,a total of 46 wheat landraces and 34 wheat lines with known Lr(leaf rust resistance)genes were inoculated with 16Pt pathotypes for postulating seedling resistance gene(s)in the greenhouse.These cultivars and five wheat differential lines with adult plant resistance(APR)genes(Lr12,Lr22b,Lr34,Lr35 and Lr37)were also evaluated for identification of slow rusting resistance in the field trials in Baoding,Hebei Province of China in the 2014–2015 and 2015–2016 cropping seasons.Furthermore,10 functional molecular markers closely linked to 10 known Lr genes were used to detect all the wheat genotypes.Results showed that most of the landraces were susceptible to most of the Pt pathotypes at seedling stage.Nonetheless,Lr1 was detected only in Hongtangliangmai.The field experimental test of the two environments showed that 38 landraces showed slow rusting resistance.Seven cultivars possessed Lr34 but none of the landraces contained Lr37 and Lr46.Lr genes namely,Lr9,Lr19,Lr24,Lr28,Lr29,Lr47,Lr51 and Lr53 were effective at the whole plant stage.Lr18,Lr36 and Lr45 had lost resistance to part of pathotypes at the seedling stage but showed high resistance at the adult plant stage.Lr34 as a slowing rusting gene showed good resistance in the field.Four race-specific APR genes Lr12,Lr13,Lr35 and Lr37 conferred good resistance in the field experiments.Seven race-specific genes,Lr2b,Lr2c,Lr11,Lr16,Lr26,Lr33 and LrB had lost resistance.The 38 landraces showed slow rusting resistance to wheat leaf rust can be used as resistance resources for wheat resistance breeding in China.

Effects of the Fhb1 gene on Fusarium head blight resistance and agronomic traits of winter wheat

The gene Fhb1 has been used in many countries to improve wheat Fusarium head blight(FHB) resistance. To make better use of this gene in the Yellow-Huai River Valleys Winter Wheat Zone(YHWZ), the most important wheat-producing region of China, it is desirable to elucidate its effects on FHB resistance and agronomic traits in different genetic backgrounds. Based on a diagnostic marker for Fhb1, six BC2 populations were developed by crossing dwarf-male-sterile(DMS)-Zhoumai 16 to three Fhb1 donors(Ningmai 9, Ningmai 13, and Jianyang 84) and backcrossing to Zhoumai 16 and Zhoumai16’s derivative cultivars(Lunxuan 136 and Lunxuan 13) using marker-assisted backcross breeding. The progenies were assessed for FHB resistance and major agronomic traits.The Fhb1 alleles were identified using the gene-specific molecular marker. The plants with the Fhb1-resistant genotype(Fhb1-R) in these populations showed significantly fewer infected spikelets than those with the Fhb1-susceptible genotype(Fhb1-S). When Lunxuan 136 was used as the recurrent parent, Fhb1-R plants showed significantly fewer infected spikelets per spike than Fhb1-R plants produced using Lunxuan 13 as the recurrent parent, indicating that the genetic backgrounds of Fhb1 influence the expression of FHB resistance. Fhb1-R plants from the DMS-Zhoumai 16/Ningmai 9//Zhoumai 16/3/Lunxuan 136 population showed the highest FHB resistance among the six populations and a significantly higher level of FHB resistance than the moderately susceptible control Huaimai 20. No significant phenotypic differences between Fhb1-R and Fhb1-S plants were observed for the eight agronomic traits investigated. These results suggest that it is feasible to improve FHB resistance of winter wheat withoutreducing yield potential by introgressing Fhb1 resistance allele into FHB-susceptible cultivars in the YHWZ.

Enhanced Stem Nematode Resistance of Transgenic Sweetpotato Plants Expressing Oryzacystatin-I Gene

Enhanced stem nematode resistance of transgenic sweetpotato （cv. Lizixiang） was achieved using Oryzacystatin-I （OCI） gene with Agrobacterium tumefaciens-mediated transformation. A. tumefaciens strain EHA105 harbors a binary vector pCAMBIA1301 with OCI gene, gusA gene and hptII gene. Selection culture was conducted using 25 mg L-1 hygromycin. A total of 1 715 plants were produced from the inoculated 1 450 cell aggregates of Lizixiang via somatic embryogenesis. GUS assay and PCR analysis of the putative transgenic plants randomly sampled showed that 90.54% of them were transgenic plants. Transgenic plants exhibited significantly enhanced resistance to stem nematodes compared to the untransformed control plants by the field evaluation with stem nematodes. Stable integration of the OCI gene into the genome of resistant transgenic plants was confirmed by Southern blot analysis, and the copy number of integrated OCI gene ranged from 1 to 4. Transgene overexpression in stem nematode-resistant plants was demonstrated by quantitative real-time PCR analysis. This study provides a way for improving stem nematode resistance in sweetpotato.

Breeding of Selectable Marker-Free Transgenic Rice Lines Containing AP1 Gene with Enhanced Disease Resistance

In order to obtain marker-free transgenic rice with improved disease resistance, the AP1 gene of Capsicum annuum and hygromycin-resistance gene （HPT） were cloned into the two separate T-DNA regions of the binary vector pSB130, respectively, and introduced into the calli derived from the immature seeds of two elite japonica rice varieties, Guangling Xiangjing and Wuxiangjing 9, mediated by Agrobacterium-mediated transformation. Many cotransgenic rice lines containing both the AP1 gene and the marker gene were regenerated and the integration of both transgenes in the transgenic rice plants was confirmed by either PCR or Southern blotting technique. Several selectable marker-free transgenic rice plants were subsequently obtained from the progeny of the cotransformants, and confirmed by both PCR and Southern blotting analysis. These transgenic rice lines were tested in the field and their resistance to disease was carefully investigated, the results showed that after inoculation the resistance to either bacterial blight or sheath blight of the selected transgenic lines was improved when compared with those of wild type.

Tagging and Utilization Bruchid Resistance Gene Using PCR Markers in Mungbean

Sixteen mungbean lines were analyzed using 56 random primers. Different DNA bands were detected between Bruchid resistant lines and susceptible lines. According to the cluster results, the 16 lines can be divided into four groups, including brucid resistant wild types, resistant cultivated lines, resistant progenies and a mixed group. BSA method was used to identify DNA markers that related with bruchid resistant gene by using resistant line and susceptible line and their F2 progeny. One codominant marker was identified, which generated a fragment of 1.79 kb in resistant lines and 1.03 kb in susceptible lines. Finally, this codominant marker was considered to be tightly linked with bruchid resistant gene and could be useful in resistant germplasm identification and marker-assisted selection.

Insulin resistance,adipokine profile and hepatic expression of SOCS-3 gene in chronic hepatitis C

AIM:to analyze adipokine concentrations,insulin resistance and hepatic expression of suppressor of cytokine signaling 3(SOCS-3)in patients with chronic hepatitis C genotype 1 with normal body weight,glucose and lipid profile.METHODS:The study group consisted of 31 patients with chronic hepatitis C and 9 healthy subjects.Total levels of adiponectin,leptin,resistin,visfatin,omentin,osteopontin and insulin were measured using an ELISA kit.The hepatic expression of SOCS-3 was determined by the use of the reverse transcription polymerase chain reaction method.RESULTS:Homeostasis model assessment for insulin resistance(HOMA-IR)values were significantly higher in hepatitis C virus(HCV)infected patients without metabolic disorders compared to healthy controls(2.24 vs 0.59,P=0.0003).Hepatic steatosis was observed in 32.2%of patients with HCV infection and was found in patients with increased HOMA-IR index(2.81 vs1.99,P=0.05)and reduced adiponectin level(5.96vs 8.37,P=0.04).Inflammatory activity(G≥2)was related to increased osteopontin concentration(34.04vs 23.35,P=0.03).Advanced liver fibrosis(S≥2)was associated with increased levels of omentin and osteopontin(436.94 vs 360.09,P=0.03 and 32.84 vs20.29,P=0.03)and reduced resistin concentration(1.40 vs 1.74,P=0.047).No correlations were reported between adipokine profile,HOMA-IR values and hepatic expression of the SOCS-3 gene.CONCLUSION:We speculated that no relationship between adipokines and HOMA-IR values may indicate that HCV can induce insulin resistance itself.Some adipokines appear to be biochemical markers of steatosis,inflammation and fibrosis in patients with chronic HCV infection.

Development of Hybrid Rice Variety FY7206 with Blast Resistance Gene Pid3 and Cold Tolerance Gene Ctb1

Hybrid rice Fanyou 7206（FY7206）, derived from the cross between a sterile line Fanyuan A and a restorer line Fuhui 7206, was bred by the Rice Research Institute, Fujian Academy of Agricultural Sciences, China. FY7206 was characterized by moderate blast resistance, cold tolerance, as well as wide adaptability, and high yields. The blast resistance results indicated that the frequencies of blast races in race B, race C and the total resistance frequency for FY7206 were 95.5%, 100.0% and 97.2%, respectively. The disease resistance results showed that the leaf blast grade for FY7206 was level 1 and panicle blast was level 5. The indoor spray results indicated that FY7206 was resistant to 11 isolates of Magnorpathe oryzae. The blast resistance of FY7206 might be derived from the high expression of blast resistance gene Pid3. The results for simulated cold resistance in an artificial climate chamber indicated that the cold tolerance for FY7206 was moderate at the booting and flowering stages. The cold tolerance results also indicated that FY7206 could be tolerant to temperatures as low as 10 °C at the seedling stage. The q RT-PCR results showed that the expression of cold tolerance gene Ctb1 in FY7206 was relatively high. These results suggested that FY7206 is a hybrid indica rice variety with good comprehensive characteristics, including blast resistance and cold tolerance.

Identification and Cloning of Resistance Gene Analogues (RGAs) Encoding NBS-LRR Proteins from Gossypium arboreum L.

Plants have developed a complicated defense mechanism during evolution to resist the harmful pathogens they encountered.The mechanism involves the interaction of the plant resistance(R)

Isolation and Functional Characterization of a B3 Transcription Factor Gene <i>FUSCA3</i>Involved in Pre-Harvest Sprouting Resistance in Wheat

Pre-harvest sprouting (PHS) reduces yields and grain quality, resulting in seriously economic losses in wheat. It has been showed that PHS is significantly correlated to seed dormancy levels. FUSCA3 (FUS3) gene is considered to be the key regulator of seed dormancy. However, little information is available about the function of FUS3 gene (TaFUS3) in wheat. In this study, three homologous genes were identified in wheat grain, and their functions were investigated by gene silencing. Three full-length DNA (3477, 3534 and 3501 bp) and cDNA (1015, 1012 and 1015 bp) sequences encoding a B3 transcription factor, designated TaFUS3-3A, TaFUS3-3B and TaFUS3-3D, were first isolated from common wheat. The transcription of three TaFUS3 genes in seed development and germination process was detected. TaFUS3-3B and TaFUS3-3D had similar expression profiles, and high levels of gene transcripts were detected in seeds at 25 DAP (days after pollination) and after 24 h of imbibition. However, the transcription of TaFUS3-3A was not detected. Silencing of TaFUS3 in common wheat spikes resulted in increased seed germination and PHS. Compared with wild-type, the TaFUS3-silenced plants showed increased expression of genes related to GA biosynthesis and ABA metabolism, and decreased expression of genes associated with ABA biosynthesis. Moreover, silencing of TaFUS3 in wheat plants led to a decrease in embryo sensitivity to ABA and changed the expression of genes involved in ABA signal transduction. The results of gene silencing indicated that TaFUS3 plays a positive role in wheat seed dormancy and PHS-resistance, which might be associated with ABA, GA level and signal transduction.

Conventional Breeding and Molecular Markers for Blast Disease Resistance in Rice (Oryza sativa L.)

Monogenic lines,which carried 23 genes for blast resistance were tested and used donors to transfer resistance genes by crossing method.The results under blast nursery revealed that 9 genes from 23 genes were susceptible to highly susceptible under the three locations(Sakha,Gemmeza,and Zarzoura in Egypt);Pia,Pik,Pik-p,Piz-t,Pita,Pi b,Pi,Pi 19 and Pi 20.While,the genes Pii,Pik-s,Pik-h,Pi z,Piz-5,Pi sh,Pi 3,Pi 1,Pi 5,Pi 7,Pi 9,Pi 12,Pikm and Pita-2 were highly resistant at the same locations.Clustering analysis confirmed the results,which divided into two groups;the first one included all the susceptible genes,while the second one included the resistance genes.In the greenhouse test,the reaction pattern of five races produced 100%resistance under artificial inoculation with eight genes showing complete resistance to all isolates.The completely resistant genes:Pii,Pik-s,Piz,Piz-5(=bi2)(t),Pita(=Pi4)(t),Pita,Pi b and Pi1 as well as clustering analysis confirmed the results.In the F1 crosses,the results showed all the 25 crosses were resistant for leaf blast disease under field conditions.While,the results in F2 population showed seven crosses with segregation ratio of 15(R):1(S),two cross gave segregated ratio of 3 R:1 S and one gave 13:3.For the identification of blast resistance genes in the parental lines,the marker K3959,linked to Pik-s gene and the variety IRBLKS-F5 carry this gene,which was from the monogenic line.The results showed that four genotypes;Sakha 105,Sakha 103,Sakha 106 and IRBLKS-F5 were carrying Pik-s gene,while was absent in the Sakha 101,Sakha 104,IRBL5-M,IRBL9-W,IRBLTACP1 and IRBL9-W(R)genotypes.As for Pi 5 gene,the results showed that it was present in Sakha 103 and Sakha 104 varieties and absent in the rest of the genotypes.In addition,Pita-Pita-2 gene was found in the three Egyptian genotypes(Sakha 105,Sakha 101 and Sakha 104)plus IRBLTACP1 monogenetic.In F2 generation,six populations were used to study the inheritance of blast resistance and specific primers to confirm the ratio and identify the resistance genes.However,the ratios in molecular markers were the same of the ratio under field evaluation in the most population studies.These findings would facilitate in breeding programs for gene pyramiding and gene accumulation to produce durable resistance for blast using those genotypes.

Identification of a RAPD marker linked to a blast resistance gene in Oryza sativa L

Marker-aided selection has received more attention in recent years. This relies on the exploitation of dose linkage between molecular markers and target gene(s). We report here a randomly amplified polymorphic DNA (RAPD) marker tightly linked to the blast resistance gene Pi-ll(t) derived from Hongjiaozhan, which confers the resistance to race ZB1 of Pyricularia oryzae Cav.

Transcriptome and Metabolome Revealed the Mechanism of NtBRL3 Overexpression Tobacco(Nicotiana tabacum L.K326)in Response to Drought Stress

Drought has severely affected the yield and quality of commercial crops.The BRI1 family plays an important role in plant response to drought stress,and BRL3 gene plays an important role in the study of drought in Arabidopsis thaliana.In this study,NtBRL3 was constructed as a vector and genetically transformed to obtain‘N.Tobacco K326’overexpression of NtBRL3.The enzyme activities of transgenic tobacco and wild-type tobacco were measured and transcriptome and metabolome analyses were performed.The results showed that the antioxidant enzymes of transgenic tobacco were more active under drought conditions,and 85 significantly differentially metabolites and 106 significantly differentially expressed genes were identified in the metabolome and transcriptome analyses,respectively.Transgenic tobacco NtBRL3ox demonstrated an excessive accumulation of droughtrelated metabolites,sugars such as sucrose and maltotetraose,and amino acids such as proline,compared with WT.We discovered drought-related differential genes in the root transcriptome,among which LOX6,RD22,WSD1,CCD8,and UGT were key genes which play an important role in plant response to drought stress.Our results demonstrate that NtBRL3 overexpression in K326 enhances drought resistance in transgenic tobacco.

基因沉默番木瓜环斑病毒复制酶基因(PRSV-Nib)获得抗病毒病番木瓜的研究

番木瓜是重要的热带经济水果。番木瓜环斑病毒(Papaya ringspot virus,PRSV)是番木瓜的重要病毒病,经常导致严重的产量损失和质量恶化。自从1998年第一例转基因番木瓜问世以来,使得基于“致病菌衍生的抗病性(pathogen-derived resistance,PDR)”的抗病育种策略获得成功广泛应用。然而依赖于序列同源性的抗病性与病毒突变导致多样性增加之间的矛盾成为番木瓜育种科学家的新挑战。本研究拟采用RNAi策略针对复制酶(nuclear inclusion b.Nib)获得广谱抗PRSV番木瓜新种质。通过团队已建立的胚性愈伤诱导-农杆菌介导转化-再生苗诱导的番木瓜遗传转化体系,共获得经过抗性筛选的再生苗52株,通过特异性PCR进行筛选共计获得24株转基因阳性植株。通过对T0代田间自然发病试验中,转基因番木瓜株系抗病性明显高于非转基因对照,其中NibB5-2田间抗病性最优。通过hiTAIL-PCR方法确定NibB5-2插入位点位于第2号染色体supercontig_30的1976766的位置。T1代接种试验中,无病毒积累且无发病症状,初步确认具有良好的病毒抗性,为番木瓜抗病育种提供新思路。

源于Halomonas sp.抗草甘膦EPSPS的克隆、鉴定及应用

为培育高抗草甘膦作物以应对草甘膦杂草进化,从海洋细菌中筛选到1株高抗草甘膦的盐单胞菌属菌株(Halomonassp.),通过基因组测序及生物信息学分析,确定该菌株的EPSPS基因,在Escherichiacoli(DE3)中对fHoEPSPS、mfHoEPSPS(G384A位点突变)和mHoEPSPS(mfHoEPSPS N端缺失PDT)进行重组表达和纯化,并运用自切割肽LP4/2A介导的基因聚合策略,将抗草铵膦的酶(Repat)置于mHoEPSPS的N端,构建了双抗草甘/铵膦酶(RLH),将其编码基因导入烟草后赋予草甘/铵膦复合抗性。结果显示,该菌株的EPSPS基因(fHoEPSPS)可编码一个N段融合了预苯酸脱水酶(PDT)的双功能酶。草甘膦抗性分析显示mfHoEPSPS的抗性比fHoEPSPS提高了19倍,将mHoEPSPS基因导入烟草后可赋予烟草3倍推荐剂量的草甘膦耐受性,转RLH基因的烟草能够耐受3~5倍推荐剂量的草甘/铵膦复合除草剂。结果表明,源于Halomonas sp.的抗草甘膦EPSPS是一种新型草甘膦耐受酶,通过G384A的位点突变可提高酶活;利用自切割肽介导的基因堆叠策略获得的转RLH基因烟草表现出较高草甘/铵膦复合抗性。

黄花苜蓿MfMYB3R-3基因克隆及结构与功能的生物信息学分析

MYB转录因子是植物中分布最广的转录因子家族之一,在植物响应生物及非生物胁迫,调控逆境胁迫相关基因表达方面起着重要的作用。本研究基于黄花苜蓿转录组数据,鉴定和克隆了黄花苜蓿MfMYB3R-3基因,并进行了生物信息学分析。结果表明,MfMYB3R-3基因CDS区长1701 bp,编码566个氨基酸,具有SANT保守结构域,为MYB3R型转录因子;MfMYB3R-3蛋白化学分子式为C_(2690)H_(4314)N_(796)O_(873)S_(20),理论等电点为8.75,该蛋白是亲水性不稳定偏碱性非分泌蛋白,不具有信号肽,无跨膜结构,亚细胞定位预测在细胞核。系统进化分析发现,黄花苜蓿MfMYB3R-3蛋白与蒺藜苜蓿的MYB3R-3的亲缘关系最近。

菊芋14-3-3基因家族的鉴定及其对非生物胁迫响应的分析

[目的]本文旨在鉴定菊芋14-3-3基因家族成员并分析它们对高温、低温、盐和干旱胁迫响应的表达模式,为研究14-3-3蛋白功能及菊芋育种提供依据。[方法]采用克隆和生物信息学研究基因性质,采用RNA-seq数据分析和RT-qPCR研究基因对非生物胁迫的响应模式。[结果]从菊芋中克隆到14-3-3基因家族的10个成员HtGRF1—HtGRF10,GenBank登录号为OP132618—OP132627。根据进化关系将其分为2个亚家族:HtGRF1—HtGRF7属于非ε组,HtGRF8—HtGRF10属于ε组,通常形成同源或异源二聚体。组织表达分析表明,HtGRF2/3/6/9在芽、根、茎、叶中的表达丰度较高,HtGRF3/5/9在块茎发育过程中前高后低。综合分析根和叶中HtGRF对非生物胁迫响应时发现,HtGRF6表达水平在高温、盐和干旱胁迫下下降;HtGRF2/3/7/8/9表达水平在盐胁迫下下降,而在干旱胁迫下上升;HtGRF1表达水平在低温胁迫下上升;HtGRF4/5表达水平在干旱胁迫下下降;而HtGRF10表达水平变化不显著。[结论]菊芋14-3-3蛋白是一个高度保守的多基因编码家族,在菊芋的生长发育和适应复杂环境中起着重要作用。

金鱼草WAK/WAKL基因家族鉴定及抗核盘菌候选基因挖掘

【目的】鉴定金鱼草细胞壁相关受体激酶(Wall-associated kinase,WAK)及类WAK蛋白(WAK-like kinases,WAKL)基因家族成员,并挖掘抗核盘菌的候选基因,为深入解析金鱼草抗核盘菌的WAK/WAKL分子调控机制提供理论参考。【方法】以拟南芥WAK/WAKL家族蛋白为参考序列,利用生物信息学方法从金鱼草基因组数据库中鉴定出WAK/WAKL基因家族成员,并对其理化性质、系统发育、基因结构、保守基序、顺式作用元件及共线性关系等进行预测分析,并通过转录组测序和实时荧光定量PCR对该家族基因在金鱼草抗感核盘菌材料中的表达模式进行分析。【结果】从金鱼草基因组中共鉴定出27个WAK/WAKL基因家族成员,其中WAK家族基因16个(AmWAK1~AmWAK16),WAKL家族基因11个(AmWAKL1~AmWAKL11),编码的氨基酸数量为615~1085个;理论等电点(pI)为4.94~8.69,绝大多数蛋白呈酸性;所有蛋白的总平均亲水性指数(GRAVY)值小于0,均为亲水性蛋白;大多数蛋白亚细胞定位于细胞质膜区域,少部分定位于细胞外、液泡、高尔基体和细胞核。27个WAK/WAKL基因家族成员不均匀地分布在金鱼草的8条染色体上,且与拟南芥WAK/WAKL家族基因存在7对共线性同源基因;基因启动子区域含有与防御和胁迫响应、茉莉酸甲酯响应及水杨酸响应等顺式作用元件。通过叶片离体接种核盘菌试验,筛选出抗性差异明显的易感病品种Am1和抗病品种Am6。有12个WAK/WAKL家族基因在金鱼草抗感材料中显著差异表达(P<0.05),有9个基因表达模式与转录组测序结果基本一致。这9个基因中,有5个基因(AmWAK7、AmWAKL2、AmWAKL8、AmWAK6和AmWAK13)在金鱼草的根、茎和叶组织中高表达,而剩余4个基因(AmWAK2、AmWAK8、AmWAK9和AmWAK12)在根、茎和叶中基本不表达。【结论】筛选出的27个金鱼草WAK/WAKL基因家族成员,具有相似的基因结构和蛋白功能结构域,其中AmWAK6、AmWAK7、AmWAK13、AmWAKL2和AmWAKL8为金鱼草抗核盘菌的关键候选基因,具有介导抗病的潜在功能。

芝麻苗期干旱生理生化指标响应特征及其基因表达分析

为建立芝麻抗旱性快速鉴定体系及筛选芝麻抗旱品种,采用盆栽反复干旱法,设置正常水分(CK)和干旱胁迫(DS)两种处理,对31份芝麻材料进行生理生化指标测定和综合评价,同时利用qRT-PCR检测SOD合成相关基因的表达量。结果表明:芝麻苗期■、可溶性糖(SS)、脯氨酸(Pro)含量和超氧化物歧化酶(SOD)、过氧化物酶(POD)活性与CK相比均显著上升,各指标综合抗旱系数和抗旱指数的变异系数最高的分别是SOD(98.64%)和■(154.01%);抗旱指数与■含量、SOD活性呈极显著正相关关系,这两个指标可作为芝麻苗期抗旱性鉴定的重要指标。聚类分析将31份芝麻材料划分为5类抗旱类型,分别为高抗型、中抗型、低抗型、低感型和高感型。利用综合评价方法,筛选出高抗材料2份(‘汾芝10号’和‘豫-11-1’),中抗旱材料4份,低抗旱材料9份,敏感材料10份,高感材料5份;筛选出■含量和SOD活性可作为芝麻种质资源苗期抗旱特性快速鉴定的指标。

Study on Mutations in ALS for Resistance to Tribenuron-Methyl in Galium aparine L.

In recent years, Galium aparine L. has not been controlled by tribenuron-methyl in major Chinese winter wheat fields. The objective of this study is to understand the molecular basis of the resistance mechanism to tribenuron-methyl in G. aparine and to find the specific mutation sites in amino acid sequence of acetolactate synthase （ALS） in the resistant biotype of G. aparine. Fragments that encode the ALS were amplified and cloned from susceptible （S） and resistant （R） biotypes of G. aparine to tribenuron-methyl and sequenced subsequently. The result showed that the nucleotide sequence of Rbiotype of G. aparine differed from that of the S biotype with three amino acid substitutions, of which, the amino acid substitution of Trp57-4 （TGG） to Gly （GGG） is located in the highly conserved region Domain B. The substitution of Trp574 might be responsible for the resistance to tribenuron-methyl in the R-biotype of G. aparine.