Identification of a target‐site mutation conferring resistance to triazine herbicides in oriental mustard (Sisymbrium orientale L.) from Australia

In southern Australia, oriental mustard (Sisymbrium orientale) has been controlled successfully by triazine herbicides for several decades. The screening of 40 populations that were collected from the southern grain belt of Australia during 2010 and 2013 for resistance to six different herbicides (glyphosate, diflufenican, imazamox, chlorsulfuron, atrazine and 2,4‐dichlorophenoxyacetic acid) identified two oriental mustard populations as highly resistant to atrazine. Compared to the known oriental mustard‐susceptible populations (S1 and S2), these two resistant populations (P17 and P18) from near Horsham, Victoria, Australia, were 311‐ and 315‐fold resistant to atrazine, as determined by a comparison of the LD₅₀ values. However, there was no resistance to diuron detected in these populations. Sequencing of the chloroplast psbA gene identified a missense mutation of serine 264 to glycine in both herbicide‐resistant oriental mustard populations, which is known to confer high‐level atrazine resistance in other species.     

A psbA mutation in Kochia scoparia (L) Schrad from railroad rights-of-way with resistance to diuron, tebuthiuron and metribuzin

Kochia [Kochia scoparia (L) Schrad] has become resistant to many herbicides used in cropland and railroad rights-of-way in North Dakota and Minnesota. Kochia scoparia plants that had survived annual treatments with diuron and tebuthiuron were sampled along railroad rights-of-way in North Dakota and Minnesota. The samples were screened in the greenhouse for resistance to diuron, tebuthiuron, metribuzin and bromoxynil from 0.5x to 32x the recommended use rates. A resistant K scoparia accession (MN-3R) was confirmed with resistance up to 16-fold higher than recommended use rates for tebuthiuron and diuron and up to 4-fold higher for metribuzin. However, the resistant K scoparia accession was susceptible to bromoxynil even at 50% of the recommended use rate. The herbicide binding region of the psbA gene fragment of eight resistant (R) and seven susceptible (S) K scoparia accessions was PCR-amplified and sequenced for detection of mutations. The psbA gene of four R K scoparia accessions was mutated at residue 219 with substitution of isoleucine for valine (GenBank accession number AY251265). The seven S K scoparia accession sequences were wild-type at this residue (GenBank accession number AY251266). The other four R accessions sequences showed a previously known triazine R mutation with substitution of glycine for serine at residue 264. All 15 K scoparia accessions were wild-type at all other psbA residues within the region analyzed. Resistance to diuron, tebuthiuron and metribuzin among the railroad rights-of-way K scoparia is probably due to the mutation at residue 219 of the psbA gene in some plants, but due to the previously reported Ser(264)Gly substitution in other plants. Target-site resistance associated with a change of valine to isoleucine at residue 219 of the psbA target-site in weeds has previously been reported for Poa annua L selected in diuron-treated grass seed fields, and for Amaranthus powelli S Wats selected in linuron-treated carrot fields. This is the first report of the mutation in herbicide-resistant K scoparia.     

紫花苜蓿下胚轴愈伤组织诱导及再生植株的研究

以培养4～5d的紫花苜蓿无菌苗的下胚轴为材料,改良SH(SH大量元素+MS微量元素+MS铁盐+UM有机)+水解酪蛋白(CH)2000 mg/L为基本培养基,对紫花苜蓿愈伤组织诱导和植株再生进行了研究。愈伤组织诱导和继代培养基分别为2,4-D 1.0 mg/L+KT 0.5 mg/L+蔗糖30 g/L及2,4-D 0.05 mg/L+BA0.5 mg/L+蔗糖30 g/L,将获得的愈伤组织转入正交设计的培养基上进行筛选,得到了愈伤组织分化率相对较高的培养基配方:基本培养基+KT 0.4 mg/L+蔗糖20 g/L,其分化率为73.0%。获得的再生芽在生根培养基上(1/2MSO)培养约10d便能获得具有根和叶的再生植株。本试验结果为下一步将来自蓝藻的Na⁺/H⁺ an-tiporter基因转入苜蓿,获得转基因耐盐苜蓿奠定了基础。     

田间小菜蛾抗药性监测及毒理机制研究

田间监测结果表明，福州地区小菜蛾Plutella xylostella(L.)已对生产上常用的各类杀虫剂产生不同程度的抗性，从9月至次年5年期间，田间小菜蛾表现出较高水平的抗性，而在8月份则表现出明显低的抗性水平，短期高温和长期低温的环境条件可明显抑制小菜蛾幼虫的AChE和CarE活性，但短期低温的环境条件对小菜蛾幼虫的AChE和CarE活性影响不大，饲养温度的变化不会影响小菜蛾幼虫AChE和Ki值，但在较高的酶与杀虫剂的反应温度下，有机磷对小菜蛾、菜青虫和黄曲条跳甲AChE的抑制作用显著增高，此外，探讨了田间影响小菜蛾抗性水平的环境因素及毒理学机制。     

Cross-resistance and inheritance of resistance to Bacillus thuringiensis toxin Cry1Ac in diamondback moth (Plutella xylostella L) from lowland Malaysia

A field population of Plutella xylostella from Malaysia (SERD4) was divided into five sub-populations and four were selected (G2-G5) with the Bacillus thuringiensis insecticidal crystal (Cry) toxins Cry1Ac, Cry1Ab, Cry1Ca and Cry1Da. Bioassay at G6 gave resistance ratios of 88, 5, 2 and 3 for Cry1Ac, Cry1Ab, Cry1Ca and Cry1Da respectively compared with the unselected sub-population (UNSEL-SERD4). The Cry1Ac-selected population showed little cross-resistance to Cry1Ab, Cry1Ca and Cry1Da, (3-, 2- and 3-fold compared with UNSEL-SERD4), whereas the Cry1Ab-SEL sub-population showed marked cross-resistance to Cry1Ac (40-fold), much greater than Cry1Ab itself. In contrast, the Cry1Ca- and Cry1Da-SEL sub-population showed little if any cross-resistance to Cry1Ac and Cry1Ab. The mode of inheritance of resistance to Cry1Ac was examined in Cry1Ac-selected SERD4 by standard reciprocal crosses and back-crosses using a laboratory insecticide-susceptible population (ROTH). Logit regression analysis of F1 reciprocal crosses indicated that resistance to Cry1Ac was inherited as an incompletely dominant trait. At the highest dose of Cry1Ac tested, resistance was recessive, while at the lowest dose it was almost completely dominant. The F2 progeny from a back-cross of F1 progeny with ROTH were tested with a concentration of Cry1Ac that would kill 100% of ROTH. The mortality ranged between 50 and 95% in seven families of back-cross progeny, which indicated that more than one allele on separate loci were responsible for resistance to Cry1Ac.     

Development of near-isogenic lines and identification of markers linked to auxinic herbicide resistance in wild mustard (Sinapis arvensis L.)

BACKGROUND: Auxinic herbicides are widely used for selective control of many broadleaf weeds, e.g. wild mustard. An auxinic‐herbicide‐resistant wild mustard biotype may offer an excellent model system to elucidate the mechanism of action of these herbicides. Classical genetic analyses demonstrate that the wild mustard auxinic herbicide resistance is determined by a single dominant gene. Availability of near‐isogenic lines (NILs) of wild mustard with auxinic herbicide resistance (R) and herbicide susceptibility (S) will help to study the fitness penalty as well as the precise characterization of this gene. RESULTS: Eight generations of backcrosses were performed, and homozygous auxinic‐herbicide‐resistant and auxinic‐herbicide‐susceptible NILs were identified from BC₈F₃ families. S plants produced significantly more biomass and seed compared with R plants, suggesting that wild mustard auxinic herbicide resistance may result in fitness reduction. It was also found that the serrated margin of the first true leaf was closely linked to auxinic herbicide resistance. Using the introgressed progeny, molecular markers linked to auxinic herbicide resistance were identified, and a genetic map was constructed. CONCLUSION: The fitness penalty associated with the auxinic herbicide resistance gene may explain the relatively slow occurrence and spread of auxinic‐herbicide‐resistant weeds. The detection of the closely linked markers should hasten the identification and characterization of this gene. Copyright © 2012 Society of Chemical Industry     

The sodium channel gene in Tetranychus cinnabarinus (Boisduval): identification and expression analysis of a mutation associated with pyrethroid resistance

BACKGROUND: The carmine spider mite (CSM), Tetranychus cinnabarinus, is the most harmful mite pest of various crops and vegetable plants. Pyrethroid insecticide fenpropathrin has been used to control insects and mites worldwide, but CSM has developed resistance to this compound. RESULTS: Three synergists together eliminated about 50% resistance against fenpropathrin in the CSM. A point mutation was identified from the sodium channel gene of fenpropathrin‐resistant CSM (FeR) by comparing cDNA sequences between FeR and susceptible (S) sodium channel genes, which caused a phenylalanine (F) to isoleucine (I) change at amino acid 1538 position in IIIS6 of the sodium channel and has been proven to confer strong resistance to pyrethroid in other species. The mRNA expression of the sodium channel gene in the FeR and abamectin‐resistant strain (AbR), which was included as a control, were both relatively lower than in the S. CONCLUSION: These results demonstrate that a mutation (F1538I) is present in the sodium channel gene in FeR of CSM, likely playing an important role in fenpropathrin resistance in T. cinnabarinus, but that decrease in the abundance of sodium channel did not confer this resistance. The F1538I mutation could be used as a molecular marker for detecting kdr resistance in Arachnida populations. Copyright © 2011 Society of Chemical Industry     

不同种群小菜蛾对4种杀虫药剂的抗性及抗性基因频率

用 FAO推荐的点滴法测定了日本、台湾、广州、东莞等地小菜蛾 (Plutella xy-lostella L .)的抗药性。测定结果表明 :日本、广州的小菜蛾对阿维菌素显示出抗药性 ,其抗性比率分别是 4 .85倍和 10 .2 7倍。杀虫药剂抗性基因型频率测定显示 ,广州、广东东莞和北京种群的最高 ,基因型频率分别为 4 1.2 5%、4 1.2 5%和 58.2 3%。研究验证了快速测定杀虫药剂抗性基因型频率的方法 ,并提出了根据不同种群小菜蛾抗性基因型频率所应采取的综合治理策略。     

白菜型油菜BraSAUR基因家族鉴定及其表达分析

SAUR(Small auxin-up RNA)是生长素早期响应逆境胁迫的基因,本文研究白菜型油菜全基因组中SAUR家族的信息,分析了该家族成员的基本特征。以强抗寒性白菜型冬油菜品种陇油7号（L7）和弱抗寒性冬油菜品种陇Lenox（X）为试验材料,采用低温和干旱处理,应用荧光定量技术分析不同基因在不同品种中的表达特性,筛选差异表达基因,为研究生长素早期应答基因调控白菜型冬油菜生长点发育机理提供支撑。结果表明：白菜型油菜的BraSAUR基因共有142个,在10条染色体中不对称分布,主要分布在2、3号染色体上,基因长度较短,且大多数基因没有内含子。qRT-PCR结果表明,低温处理后,与CK相比,在两个品种的叶片中,Bra029452的表达量变化趋势不同,24 h时在L7中的表达量是X的24倍,在生长点中,Bra010501在L7中24 h时表达量达到CK的4倍,而在X中是先增加后降低的趋势。模拟干旱胁迫后,Bra029452在L7的叶片中的表达量均显著高于CK,在X中则是逐渐降低,在L7的生长点中1 h和24 h表达量均高于CK,但在X中则表现为先降低后升高。低温胁迫和干旱胁迫条件下,Bra029452基因表达量在强抗寒性油菜品种L7的叶片及生长点中均逐渐升高,可推断该基因同时参与白菜型冬油菜耐受低温、干旱的调控过程。