ITS序列及其SNP位点在外来入侵杂草长芒苋、西部苋和糙果苋物种鉴定中的应用

对于以种子形态为物种主要识别依据的口岸部门,外来入侵杂草长芒苋、西部苋和糙果苋一直是鉴定的难点。本文对34种苋属植物的ITS序列和26SrDNA进行分析,通过SNP变异位点及特异性引物,借助PCR-RFLP方法,对长芒苋、西部苋与糙果苋进行快速、准确的分类鉴定。ITS序列分析表明,长芒苋种内变异小,种间变异显著,可与其他苋属植物明显区分。西部苋与糙果苋之间ITS序列差异小,需依据SNP位点来区别。     

糙果苋KASP检测方法的建立

检疫性杂草糙果苋（Amaranthus tuberculatus）与同属其他种的鉴别一直是个难点。本研究建立糙果苋的KASP(Kompetitive allele-specific PCR)检测方法,可用于糙果苋快速、准确的分子鉴定。提取11种23份苋属植物的DNA后,根据长芒苋（Amaranthus palmeri）核基因组序列,设计引物AP4F和AP4R,通过序列扩增和比对,发现糙果苋有特异性SNP (Single nucleotide polymorphism)位点。利用引物设计软件Primer 5设计了用于鉴定糙果苋的KASP反应引物。结果表明,特异引物基于KASP技术扩增后,可以形成明显的基因分型,能将糙果苋与同属近缘种进行区分。该研究为进口粮谷中糙果苋的检测鉴定提供了新技术。     

进境粮谷中携带苋属检疫性杂草籽风传播扩散距离的预测

风媒传播是外来杂草传播扩散的主要途径之一。我国进口粮谷中经常携带大量苋属杂草种子,这些种子质量轻,在卸货、运输等过程中存在风传播扩散风险。本文以3种重要苋属检疫性杂草,即长芒苋、西部苋和糙果苋种子为材料,通过测定其在可能撒漏高度范围和撒漏点风速范围内的垂直沉降速度和水平扩散距离,预测苋属3种检疫性杂草在装卸及运输过程中风传播扩散范围。结果表明在风速为35 m/s,种子释放高度为35 m时,长芒苋、西部苋和糙果苋3种杂草种子的最远扩散距离分别为57.75 m、111.68 m和114.33 m。     

利用双重数字PCR检测进口美国大豆中长芒苋的EPSPS基因拷贝数

长芒苋是一种原产北美洲的雌雄异株杂草.在美国,一些长芒苋种群已经对草甘膦产生了抗药性.EPSPS基因拷贝数的增加是长芒苋产生抗药性的主要原因.本文建立了双重数字PCR检测长芒苋EPSPS基因拷贝数的方法,对进口美国大豆中携带的8个长芒苋样品进行检测,发现3个样品的EPSPS相对拷贝数较低,5个样品EPSPS相对拷贝数大...     

外来入侵杂草长芒苋

长芒苋(Amaranthus palmeri S.Watson)是20世纪80年代中期传人我国的一种新植物,为中国苋属异株亚属一新归化物种.近年来长芒苋在北京市不断扩展蔓延.本文对长芒苋的形态特征、生物学特性、分布、危害及其防除作一简要介绍.     

全国口岸近年来截获苋属杂草疫情分析

苋属很多种类是农田恶性杂草,我国出入境口岸近年来在进境检疫过程中经常截获苋属杂草。本文对我国口岸在2003~2014年截获的苋属杂草疫情进行了统计分析:自2003年以来,全国口岸截获苋属杂草的种类和次数逐年递增,2003~2014年8月,全国口岸共截获苋属杂草15种29291次,共来自全世界72个国家或地区。另外,本文还统计分析了几种主要进境粮谷中截获苋属杂草的种类和次数,截获检疫性苋属杂草的主要货物,并针对苋属杂草检疫存在的问题提出了几点建议。     

基于4种生态位模型的长芒苋潜在适生区预测

恶性入侵杂草长芒苋Amaranthus palmeri S.Watson对农业生产和生物多样性造成严重威胁。预测其潜在适生区对粮食安全和生物多样性保护至关重要。不同模型由于算法不同对长芒苋潜在适生区的预测结果存在差异。本研究综合4种生态位模型(MaxEnt、GARP、BIOCLIM和DOMAIN)预测长芒苋在我国的潜在适生区以提高预测准确性。结果表明,4种模型的平均AUC值均大于0.85。MaxEnt和DOMAIN的平均Kappa值大于0.81;BIOCLIM和GARP的平均Kappa值大于0.69。MaxEnt的预测精度和稳定性要略胜一筹。BIOCLIM和MaxEnt的预测结果较为收敛,DOMAIN和GARP预测的适生区范围较广,由此预测的长芒苋潜在适生区分别占我国陆地总面积的20.66%和32.38%,48.39%和49.76%。综合预测结果,长芒苋在我国的适生区主要集中在中东部地区,西北和东北地区则是长芒苋存在的边缘环境地区。     

我国首次截获检疫性有害生物——西部苋

2011年7月,江苏首次进口的美国玉米在镇江口岸靠港。镇江局检验检疫人员对该批玉米实施检疫,并取样送江苏局动植食中心检测。经检测鉴定发现,该批玉米携带西部苋(Amaranthus rudis)、长芒苋(Amaranthus palmeri)、豚草     

基于种子发芽率评估长芒苋在我国不同纬度地区的入侵风险

外来入侵植物长芒苋Amaranthus palmeri已经传入我国多个地区,定量评估其入侵风险对制定高效防控措施具有重要意义。外来植物入侵风险应该是由传入后能发芽并完成生活史的繁殖体如种子的量决定的,但已开展的适生区预测等风险评估通常忽略了该因素。基于此,本研究先通过同质园试验, 比较了采自我国不同纬度的11个长芒苋种群与来自原产地美国种群的种子发芽率,分析了种群间发芽率的差异及其与纬度的相关性;然后,通过一年内连续多次在不同纬度地区的交互种植试验,分析了不同纬度种群间种子的发芽率和完成生活史的入侵窗口期差异,并判断是否发生了本地适应;最后,根据发芽率和入侵窗口期评估了长芒苋在我国不同纬度地区的入侵风险。同质园试验表明,种子发芽率与种群所处纬度显著正相关(P<0.05),发芽率随纬度升高而增高。交互种植试验表明,不同种群发芽率的差异是由于本地适应导致的,长芒苋在我国的入侵窗口期随纬度升高而缩短。基于不同种群种子发芽率和入侵窗口期的纬度差异,我们判断长芒苋在我国中低纬度至中高纬度区域内入侵风险较高,应该重点防控。长芒苋在高纬度和低纬度地区的入侵风险相对较低。但长芒苋种子萌发的本地适应可能会增加其在高、低纬度地区的入侵风险。因此,亟须加强对已传入种群的监测预警与早期防控力度,抑制其繁殖增长和进一步的扩散蔓延。     

靶标基因突变导致入侵性杂草长芒苋对咪唑乙烟酸产生抗性

长芒苋Amaranthus palmeri生长迅速,适应性广,繁殖系数高,具有很强的竞争性,已在我国多地定植,对作物产量及生态环境构成潜在威胁.一旦其对除草剂产生抗性,将大大增加治理难度.本试验研究了采自不同地点的长芒苋种群对除草剂咪唑乙烟酸的抗性水平和抗性机理.整株生物测定得出,长芒苋疑似抗性种群和敏感种群对咪唑乙烟...     

Triazine resistance in Amaranthus tuberculatus (Moq) Sauer that is not site-of-action mediated

While surveying Illinois Amaranthus tuberculatus (Moq) Sauer (tall waterhemp) half-sib populations for herbicide response variability, several were observed to segregate for resistance to atrazine. Studies were conducted on greenhouse-grown A tuberculatus plants to compare atrazine responses among populations that were segregating for resistance (SegR), uniformly sensitive (UniS) or uniformly resistant (UniR). In chlorophyll fluorescence assays, leaves of plants from the SegR and UniS populations displayed changes in fluorescence after treatment with atrazine, indicating that atrazine was inhibiting electron transport of photosystem II in chloroplasts. Sequencing of a fragment of psbA, which encodes the D1 protein, revealed that the SegR population did not contain the amino acid substitution that is typically found in triazine-resistant plants. Whole-plant herbicide dose-response experiments revealed that, relative to the UniS population, atrazine resistances in the UniR and SegR populations were > 770-fold and 16-fold, respectively. The SegR population was also resistant to cyanazine (59-fold), but not to metribuzin, linuron or pyridate. Triazine resistance in the SegR population was shown to be a nuclear inherited trait, unlike maternal inheritance of site-of-action mediated triazine resistance found in the UniR population. Taken collectively, these findings confirm the existence of two distinct triazine resistance mechanisms in A tuberculatus.     

Resistance to HPPD-inhibiting herbicides in a population of waterhemp (Amaranthus tuberculatus) from Illinois, United States

BACKGROUND: A population of waterhemp in a seed maize production field in central Illinois, United States, was not adequately controlled after post‐emergence applications of herbicides that inhibit 4‐hydroxyphenylpyruvate dioxygenase (HPPD). RESULTS: Progeny from the field population survived following treatment with mesotrione, tembotrione or topramezone applied to the foliage either alone or in combination with atrazine in greenhouse experiments. Dose–response experiments indicated that the level of resistance to the HPPD inhibitor mesotrione is at least tenfold relative to sensitive biotypes. CONCLUSION: These studies confirm that waterhemp has evolved resistance to HPPD‐inhibiting herbicides. Copyright © 2011 Society of Chemical Industry     

反枝苋种子休眠解除方法研究

研究了光照和黑暗两种条件下氢氧化钠（NaoH）、盐酸（HC l）、赤霉素（GA3）和乙烯利（ETH）处理对反枝苋种子萌发的影响。结果表明,氢氧化钠、盐酸、赤霉素和乙烯利浸泡处理均可以明显提高反枝苋种子的发芽率,且随着试剂浓度的提高,发芽率表现出单峰变化曲线,浓度过低或过高均不利于种子的萌发。其中,赤霉素对解除反枝苋种子休眠最有效,且以200 mg/L浸泡处理24 h效果最好,发芽率可达90.8%,比对照提高了52.5百分点。其次为4%NaOH和6%HC l处理1 h,与800 mg/L ETH处理24 h的效果相当,发芽率在70%~75%之间。黑暗条件不利于反枝苋种子的萌发,发芽率略低于光照条件。     

反枝苋对咪唑乙烟酸抗性水平及分子机制

为初步明确大豆田反枝苋对咪唑乙烟酸的抗药性水平,并从分子角度对抗药性机制进行解释,以我国四川成都和黑龙江嫩江采集的反枝苋种子为材料,通过琼脂法检测了反枝苋对咪唑乙烟酸的抗药性水平,并分别对R(嫩江抗性种群)和S(成都敏感种群)的乙酰乳酸合成酶(ALS)部分序列进行扩增和测序。皿内生测结果表明,成都种群的GI50为11.20,嫩江种群的GI50为52.26,其抗药性指数RI为4.67。分子检测结果表明,与S种群相比,R种群反枝苋ALS位于高度保守区Domain B编码574位氨基酸的基因发生突变,TGG突变为TTG,导致色氨酸被亮氨酸取代。ALS保守区域氨基酸的替换可能是嫩江反枝苋种群对咪唑乙烟酸产生抗性的重要原因之一。     

黑龙江省大豆田反枝苋对氟磺胺草醚的抗药性机制研究

为明确反枝苋抗性种群对氟磺胺草醚的抗性机制,分别测定了氟磺胺草醚对反枝苋抗性和敏感种群体内原卟啉原氧化酶(PPO)、谷胱甘肽S-转移酶(GSTs)、超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性的影响。结果表明:氟磺胺草醚处理后,抗性和敏感反枝苋种群PPO活性均受到一定程度的抑制,但抗性种群活性受到抑制后能逐渐恢复,而敏感种群则不能恢复;施用氟磺胺草醚后,抗性和敏感反枝苋种群GSTs和SOD活性变化无明显差异,抗性和敏感反枝苋种群POD和CAT活性均受到一定程度的抑制,但抗性种群活性受到抑制后能逐渐恢复,而敏感种群则不能恢复。研究表明,反枝苋抗性种群体内PPO对氟磺胺草醚敏感性降低是其产生抗药性的原因之一,反枝苋POD和CAT对活性氧的抵御能力差异也可能与反枝苋对氟磺胺草醚的抗性有关。     

Amaranthus palmeri resistance and differential tolerance of Amaranthus palmeri and Amaranthus hybridus to ALS-inhibitor herbicides

Suspected imazaquin-resistant accessions of Amaranthus palmeri were studied to determine the magnitude of resistance and cross-resistance to acetolactate synthase (ALS)-inhibiting herbicides and compare differential tolerance of A palmeri and Amaranthus hybridus to ALS inhibitors. Five of seven A palmeri accessions were resistant to imazaquin. The most imazaquin-resistant accession, accession 7, also showed 74, 39 and 117 times higher resistance than the susceptible biotype to chlorimuron, diclosulam and pyrithiobac, respectively. Resistance to imazaquin and cross-resistance to other ALS inhibitors in A palmeri was due to a less-sensitive ALS enzyme. A palmeri was 70 times more tolerant to imazaquin than A hybridus. A palmeri was also seven times more tolerant to pyrithiobac than A hybridus. Differences in ALS enzyme sensitivity could not fully account for the high tolerance of A palmeri to imazaquin compared to A hybridus. Both species were equally affected by chlorimuron and diclosulam.     

Validation of a weed:crop competition model

A model of crop:weed competition, based on parameters from the logistic biomass growth of both crop and weeds in monocultures, was validated under several sets of conditions. Model inputs were (i) dry matter per unit area at 50% of seedling emergence ( B_i ) for each species; (ii) maximum total biomass of each species in monocultures ( K ); (iii) intrinsic growth rate of each species in monocultures ( r ); (iv) the accumulated thermal time required for each species to reach 50% emergence; and (v) the total accumulated thermal time during the growing season considered in the simulation. The model adequately described the dynamics of the competition between species when the relative yield total of the mixture was close to one, i.e. the slope and intercept of the regression between observed and simulated values was not significantly different from one and zero respectively. Sensitivity analysis indicated that final crop biomass was particularly affected by changes in the relative growth rate of the species. The occurrence of a critical period for weed control was explored using the model. The simplicity of the model validated here would allow its use as a tool to predict the outcome of competition and species relative importance, particularly when parameters needed to run more complex models are not available.     

Predispersal seed predation of Amaranthus retroflexus and Chenopodium album growing in soyabean fields

Field experiments were made in 1998 and 1999 to determine the influence of tillage and soyabean (Glycine max) row width on predispersal weed seed predation in Amaranthus retroflexus L. (redroot pigweed) and Chenopodium album L. (common lambsquarters). Soyabean was planted in wide (76 cm) and narrow (19 cm) rows with conventional or conservation tillage. Additional control plots without soyabean were also established. The two objectives were to determine (1) whether predispersal seed predation occurs in A. retroflexus or C. album, and (2) whether disturbance (soil tillage) or microclimate (planting pattern) influence predation level. Mean rates of seed predation were 26% and 4% in A. retroflexus and C. album, respectively. Although these levels were low at the population level, individual plants of both species had predation levels ranging from 0% to 80%, however, very few individuals of C. album had levels of predation above 10%. Differences among tillage and row width treatments occurred for A. retroflexus, but not for C. album. Amaranthus retroflexus and C. album growing within the soyabean crop received less light than those in the no‐crop plots, and produced less above‐ground biomass, smaller terminal inflorescences, and fewer seeds per inflorescence. Plant height, terminal inflorescence weight, and total seeds were correlated with predation in both weed species.     

Sulfometuron-resistant Amaranthus retroflexus: cross-resistance and molecular basis for resistance to acetolactate synthase-inhibiting herbicides

A biotype of Amaranthus retroflexus L. is the first weed in Israel to develop resistance to acetolactate synthase (ALS)-inhibiting herbicides. The resistant biotype (Su-R) was collected from Ganot, a site that had been treated for more than 3 consecutive years with sulfometuron-methyl + simazine. On the whole-plant basis, the resistance ratio ( ED₅₀ Su-R)/( ED₅₀ Su-S) was 6–127 for sulfonylureas, 4–63 for imidazolinones, 20–35 for triazolopyrimidines and 11 for pyrithiobac-sodium. Similar levels of resistance were found also when the herbicides were applied before emergence. Based on a root elongation bioassay, Su-R was 3240-fold more resistant to sulfometuron-methyl than Su-S. In vitro studies have shown that the Su-R biotype was resistant at the enzyme level to all ALS inhibitors tested. The nucleotide sequences of two amplified regions between the Su-S and the Su-R differed in only one nucleotide. One substitution has occurred in domain A, cytosine by thymine (C C C to C T C) at position 248, that confers an exchange of the amino acid proline in the susceptible to leucine in the Su-R. The proline to leucine change in domain A is the only difference in the amino acid primary structure of the regions sequenced, indicating that it is responsible for the ALS-inhibitor resistance observed.