Physiological and biochemical characterization of quinclorac resistance in a false cleavers (Galium spurium L.) biotype

The physiological and biochemical basis for quinclorac resistance in a false cleavers (Galium spurium L.) biotype was investigated. There was no difference between herbicide resistant (R) and susceptible (S) false cleavers biotypes in response to 2,4-D, clopyralid, glyphosate, glufosinate-ammonium, or bentazon. On the basis of GR(50) (growth reduction of 50%) or LD(50) (lethal dose to 50% of tested plants) values, the R biotype was highly resistant to the acetolactate synthase (ALS) inhibitor, thifensulfuron-methyl (GR(50) resistance ratio R/S = 57), and quinolinecarboxylic acids (quinclorac R/S = 46), resistant to MCPA (R/S = 12), and moderately resistant to the auxinic herbicides picloram (R/S = 3), dicamba (R/S = 3), fluroxypyr (R/S = 3), and triclopyr (R/S = 2). The mechanism of quinclorac resistance was not due to differences in [(14)C]quinclorac absorption, translocation, root exudation, or metabolism. Seventy-two hours after root application of quinclorac, ethylene increased ca. 3-fold in S but not R plants when compared to controls, while ABA increased ca. 14-fold in S as opposed to ca. 3-fold in R plants suggesting an alteration in the auxin signal transduction pathway, or altered target site causes resistance in false cleavers. The R false cleavers biotype may be an excellent model system to further examine the auxin signal transduction pathway and the mechanism of quinclorac and auxinic herbicide action.     

Metabolism-based resistance of a wild mustard (Sinapis arvensis L.) biotype to ethametsulfuron-methyl

Under controlled-environment conditions, ethametsulfuron-methyl doses that inhibited growth by 50% (ED(50)) were >100 and <1 g of active ingredient (ai) ha(-)(1) for ethametsulfuron-methyl-resistant (R) and -susceptible (S) wild mustard, respectively. There were no differences between the two biotypes with regard to absorption and translocation of the herbicide. Three days after treatment, approximately 90, 5, and 2% of the applied [(14)C]ethametsulfuron-methyl was found in the treated leaf, foliage, and roots of each biotype, respectively. Acetolactate synthase extracted from the two biotypes was equally sensitive to both ethametsulfuron-methyl and chlorsulfuron. These results indicate that resistance was not due to differences in the target site, absorption, or translocation. However, ethametsulfuron-methyl was metabolized more rapidly in the R than the S biotype. Approximately 82, 73, 42, 30, and 17% of the recovered radioactivity remained as ethametsulfuron-methyl in R wild mustard 3, 6, 18, 48, and 72 h after treatment, respectively. Conversely, 84, 79, 85, and 73% of the (14)C was ethametsulfuron-methyl in the S biotype 12, 24, 48, and 72 h after treatment, respectively. On the basis of these results, it is proposed that resistance is due to enhanced metabolism of ethametsulfuron-methyl in the R biotype.     

Inheritance and physiological basis for 2,4-D resistance in prickly lettuce (Lactuca serriola L.)

Experiments were conducted to determine the inheritance and physiological basis for resistance to the synthetic auxinic herbicide (2,4-dichlorophenoxy)acetic acid (2,4-D) in a prickly lettuce biotype. Inheritance of 2,4-D resistance in prickly lettuce is governed by a single codominant gene. Absorption and translocation were conducted using (14)C-2,4-D applied to 2,4-D-resistant and -susceptible biotypes. At 96 h after treatment (HAT), the resistant biotype absorbed less applied 2,4-D and retained more 2,4-D in the treated portion of the leaf compared to the susceptible biotype. The resistant biotype translocated less applied 2,4-D to leaves above the treated leaf and crown at 96 HAT compared to the susceptible biotype. No difference in the rate of metabolism of 2,4-D was observed between the two biotypes. Resistance to 2,4-D appears to originate from a reduced growth deregulatory and overstimulation response compared to the susceptible biotype, resulting in lower translocation of 2,4-D in the resistant prickly lettuce biotype.     

Lolium rigidum, a pool of resistance mechanisms to ACCase inhibitor herbicides

Three diclofop-methyl (DM) resistant biotypes of Lolium rigidum (R1, R2, and R3) were found in different winter wheat fields in Spain, continuously treated with DM, DM + chlortoluron, or DM + isoproturon. Herbicide rates that inhibited shoot growth by 50% (ED50) were determined for DM. There were found that the different biotypes exhibited different ranges of resistance to this herbicide; the resistant factors were 7.2, 13, and 36.6, respectively. DM absorption, metabolism, and effects on ACCase isoforms were examined in these biotypes of L. rigidum. The most highly resistant, biotype R3, contained an altered isoform of ACCase. In biotype R2, which exhibited a medium level of resistance, there was an increased rate of oxidation of the aryl ring of diclofop, a reaction most likely catalyzed by a cytochrome P450 enzyme. In the other biotype, R1, DM penetration was significantly less than that observed in the resistant (R2 and R3) and susceptible (S) biotypes. Analysis of the leaf cuticle surface by scanning electron microscopy showed a greater epicuticular wax density in the leaf cuticles of biotype R1 than in the other biotypes.     

Late watergrass (Echinochloa phyllopogon): mechanisms involved in the resistance to fenoxaprop-p-ethyl

Fenoxaprop-p-ethyl (FE), 2-[4-[(6-chloro-2-benzoxazolyl)oxy]phenoxy] propanoate, ethyl ester (R), is an aryloxyphenoxypropionate herbicide for postemergence control of annual and perennial grasses in paddy fields; its site of action is acetyl-coenzyme A carboxylase (ACCase), an enzyme in fatty acids biosynthesis. The possible mechanism(s) of resistance to FE in a resistant biotype of Echinochloa phyllopogon was examined, namely, absorption, translocation, and metabolism of FE and ACCase susceptibility to fenoxaprop acid (FA). Studies of the in vitro inhibition of ACCase discounted any differential active site sensitivity as the basis of resistance to FE. There were differences in absorption rates between biotypes from 3 to 48 h after application (HAA). Biotypes did not differ in either the amounts or the rates of FE translocated; 98% of applied [14C]FE remaining in the treated leaf. However, there was a good correlation between the rate of herbicide metabolism and the plant resistance. The R biotype produced 5-fold less FA and approximately 2-fold more nontoxic (polar) metabolites 48 HAA than the S biotype. Moreover, the higher rate of GSH conjugation in the resistant biotype as compared to the susceptible one indicates that GSH and cysteine conjugation is the major mechanism of resistance of the R biotype against FE toxicity.     

MCPA (4-Chloro-2-ethylphenoxyacetate) resistance in hemp-nettle (Galeopsis tetrahit L.)

The physiological basis for MCPA resistance in a hemp-nettle (Galeopsis tetrahit L.) biotype, obtained from a MCPA-resistant field population, was investigated. Dose-response studies revealed that the resistance factor for MCPA, based on GR50 comparisons of total dry weight of resistant (R) and susceptible (S) plants, was 3.3. Resistance factors for fluroxypyr, dicamba, 2,4-D, glyphosate, and chlorsulfuron were 8.2, 1.7, 1.6, 0.7, and 0.6, respectively. MCPA resistance was not due to differences in absorption, because both R and S biotypes absorbed 54% of applied [14C]MCPA 72 h after treatment. However, R plants exported less (45 vs 58% S) recovered 14C out of treated leaves to the apical meristem (6 vs 13% S) and root (32 vs 38% S). In both biotypes, approximately 20% of the 14C recovered in planta was detected as MCPA metabolites. However, less of the 14C recovered in the roots of R plants was MCPA. Therefore, two different mechanisms protect R hemp-nettle from MCPA phytotoxicity: a lower rate of MCPA translocation and a higher rate of MCPA metabolism in the roots. In support of these results, genetic studies indicated that the inheritance of MCPA resistance is governed by at least two nuclear genes with additive effects.     

Mechanism of resistance to ACCase-inhibiting herbicides in wild oat (Avena fatua ) from Latin America

Whole-plant response of two suspected resistant Avena fatua biotypes from Chile and Mexico to ACCase-inhibiting herbicides [aryloxyphenoxypropionate (APP), cyclohexanedione (CHD), and pinoxaden (PPZ)] and the mechanism behind their resistance were studied. Both dose-response and ACCase enzyme activity assays revealed cross-resistance to the three herbicide families in the biotype from Chile. On the other hand, the wild oat biotype from Mexico exhibited resistance to the APP herbicides and cross-resistance to the CHD herbicides, but no resistance to PPZ. Differences in susceptibility between the two biotypes were unrelated to absorption, translocation, and metabolism of the herbicides. PCR generated fragments of the ACCase CT domain spanning the potential mutations sited in the resistant and susceptible biotypes were sequenced and compared. A point mutation was detected in the aspartic acid triplet at the amino acid position 2078 in the Chilean biotype and in isoleucine at the amino acid position 2041 in the Mexican wild oat biotype, which resulted in a glycine triplet and an asparagine triplet, respectively. On the basis of in vitro assays, the target enzyme (ACCase) in these resistant biotypes contains a herbicide-insensitive form. This is the first reported evidence of resistance to pinoxaden in A. fatua.     

Enhanced resistance to Helicoverpa zea in tobacco expressing an activated form of maize ribosome- inactivating protein

Progeny of two transgenic tobacco (Nicotiana tabacum L.) lines that expressed an activated form of maize (Zea mays L.) ribosome-inactivating protein (RIP) had varying resistance to the insect species tested. A subset of R(2) plants from the two lines appeared to be more resistant to larvae of the cigarette beetle, Lasioderma serricorne (F.), and the tobacco hornworm, Manduca sexta (L.) than the wild type plants. Progeny (R(3)) of the more resistant R(2) plants were tested more extensively for insect resistance. Resistance to the corn earworm, Helicoverpa zea (Boddie), was most consistent, with significantly decreased feeding often accompanied by increased mortality and reduced weights of survivors fed on leaf disks of the two transgenic lines compared to the wild type. The amount of damage by H. zea was significantly inversely correlated with levels of RIP. Resistance of RIP-producing plants to H. zea was greater than expected on the basis of prior in vitro results using diet-incorporated maize RIP. The R(3) transgenic plant leaf disks were also often more resistant to feeding by larvae of L. serricorne compared to wild type plants. Although reduced feeding by M. sexta was noted when they were fed leaf disks from transgenic compared to wild type plants the first day of exposure, differences were not significant. This information provides further support for maize RIP having a role in resistance to maize-feeding insects.     

Pool of resistance mechanisms to glyphosate in Digitaria insularis

Digitaria insularis biotypes resistant to glyphosate have been detected in Brazil. Studies were carried out in controlled conditions to determine the role of absorption, translocation, metabolism, and gene mutation as mechanisms of glyphosate resistance in D. insularis. The susceptible biotype absorbed at least 12% more (14)C-glyphosate up to 48 h after treatment (HAT) than resistant biotypes. High differential (14)C-glyphosate translocation was observed at 12 HAT, so that >70% of the absorbed herbicide remained in the treated leaf in resistant biotypes, whereas 42% remained in the susceptible biotype at 96 HAT. Glyphosate was degraded to aminomethylphosphonic acid (AMPA), glyoxylate, and sarcosine by >90% in resistant biotypes, whereas a small amount of herbicide (up to 11%) was degraded by the susceptible biotype up to 168 HAT. Two amino acid changes were found at positions 182 and 310 in EPSPS, consisting of a proline to threonine and a tyrosine to cysteine substitution, respectively, in resistant biotypes. Therefore, absorption, translocation, metabolism, and gene mutation play an important role in the D. insularis glyphosate resistance.     

Genetic engineering of maize (Zea mays) for high-level tolerance to treatment with the herbicide dicamba

Herbicide-tolerant crops have been widely and rapidly adopted by farmers in several countries due to enhanced weed control, lower labor and production costs, increased environmental benefits, and gains in profitability. Soon to be introduced transgenic soybean and cotton varieties tolerant to treatments with the herbicide dicamba offer prospects for excellent broadleaf weed control in these broadleaf crops. Because monocots such as maize (Zea mays) can be treated with dicamba only during a limited window of crop development and because crop injury is sometimes observed when conditions are unfavorable, transgenic maize plants have been produced and tested for higher levels of tolerance to treatment with dicamba. Maize plants expressing the gene encoding dicamba monooxygenase (DMO) linked with an upstream chloroplast transit peptide (CTP) display greatly enhanced tolerance to dicamba applied either pre-emergence or postemergence. Comparisons of DMO coupled to CTPs derived from the Rubisco small subunit from either Arabidopsis thaliana or Z. mays showed that both allowed production of transgenic maize plants tolerant to treatment with levels of dicamba (i.e., 27 kg/ha) greatly exceeding the highest recommended rate of 0.56 kg/ha.     

Bioactivity of the fungal metabolite (8R,16R)-(-)-pyrenophorin on graminaceous plants

A secondary metabolite was isolated from cultures of a Drechslera avenae pathotype with host specificity to Avena sterilis and identified as the macrodiolide (8R,16R)-(-)-pyrenophorin (8,16-dimethyl-1,9-dioxa-cyclohexadeca-3,11-diene-2,5,10,13-tetraone). A considerable yield of the substance was obtained after 8-12 days of incubation at temperatures of 15-20 degrees C. The compound at a concentration of 60 microM inhibited seed germination of wild oats (Avena sterilis, A. fatua), oat (A. sativa), wheat (Triticum aestivum), and barley (Hordeum vulgare). Root growth of pregerminated seeds of the graminaceous plants was stimulated, remained unaffected, or was inhibited by pyrenophorin at 10-30, 31-50, and >51 microM, respectively. The metabolite caused abnormal chlorophyll retention in leaf sections of all five graminaceous plants, but seedling cuttings partially immersed in 1000 microM solutions remained unaffected. The rate of chlorophyll dissipation was decreased by half in leaf sections treated with pyrenophorin at 320 microM compared with the control. These findings are discussed and compared with data on the production and bioactivity of the macrodiolide (5S,8R,13S,16R)-(-)-pyrenophorol, which has a similar stereochemical configuration.     

Germination differentiation patterns of wild and domesticated columnar cacti in a gradient of artificial selection intensity

Germination of wild and managed populations of columnar cacti was compared to analyze differences associated to management intensity. The species studied: Polaskia chende, Escontria chiotilla, Myrtillocactus schenckii, Polaskia chichipe, and Stenocereus pruinosus are in a gradient from lower to higher management intensity, respectively. Within each species wild, silvicultural, and cultivated populations also represent a gradient from lower to higher management intensity. We hypothesized that seeds of plants under higher management intensity would require more water to germinate than others since managed environments are relatively wetter than unmanaged environments. Such pattern could be visualized within and among species according to management intensity. We tested germination of seeds from different populations of the different species to the water potentials: 0.0, ?0.2, ?0.4, ?0.6, ?0.8, and ?1.0 MPa. In all species studied seeds of managed populations had significantly more mass than seeds from wild populations. Each species had different susceptibility to water availability decrease, which could be related to particular adaptations to drought resistance. Stenocereus pruinosus and Polaskia chende were the most and least susceptible species, respectively. In control treatments of all species, seeds of managed populations had higher germination rate than seeds from wild populations, but when water availability decreased the pattern was the contrary. Our results suggest that artificial selection for larger fruits and heavier seeds in plants established in wetter environments have affected germination patterns. Although species have particular critical points of water availability for germination, differences in germination patterns are more pronounced according to management and artificial selection intensity.     

狗蔷薇RaWUS基因组成型表达对烟草叶片形态的影响

为验证从狗蔷薇类原球茎克隆的RaWUS基因的功能,构建了组成型表达载体,并用根瘤农杆菌介导的叶盘转化法对烟草进行了遗传转化。通过除草剂草丁膦筛选,获得了转基因植株并对其进行了表型观察和RT．PCR分析。结果显示转基因烟草叶片形态和叶脉表现出形态变化,包括叶片浅裂、叶片边缘波浪状、叶脉紊乱,甚至叶片的主脉上有不定芽的产生等。RT-PCR分析表明,RaWUS基因仅仅在经过草丁膦筛选的抗性转基因烟草中强烈表达,在野生型烟草中没有表达。表明RaWUS基因已经被成功转入烟草中,并可能通过改变激素的水平和调控维管束的发育来影响叶片形状。     

贵州本土半野生番茄砧木介导ABA生物合成信号通路调控植株耐旱性的机理研究

随着全球气候变暖,干旱胁迫成为限制番茄等蔬菜作物安全生产的重要因素之一。前期研究发现嫁接贵州本土半野生番茄GZ-01砧木其可以提高植株的耐旱性。为探究野生番茄GZ-01增强植株耐旱性的分子机理,以半野生番茄GZ-01砧木和红果番茄为试验材料,结合形态生理学和分子生物学,比较嫁接番茄和自嫁接番茄植株对干旱胁迫的响应。结果表明,干旱胁迫下,与自嫁接植株R/R相比,GZ-01/R嫁接植株细胞膜的损伤显著降低,植株的抗氧化能力、干物质累积量、CO2同化率和水分利用率显著提高,离体叶片失水率显著降低,下气孔闭合比率显著提高,脱落酸（ABA）合成相关基因表达量和ABA含量显著提高。嫁接植株GZ-01/R可能通过调控ABA的合成来影响气孔开闭,调控叶片失水率,提高植株水分利用率,从而影响植株对干旱胁迫的响应。本研究为开发利用贵州本土半野生番茄资源奠定了基础,为增强干旱胁迫和分子调控网络途径提供了理论支撑。     

野燕麦繁殖和抗逆特性及其对小麦的他感效应研究

作为一种分布广泛的农田恶性杂草,野燕麦具有很强的环境胁迫适应能力,并且在种间竞争中常处于优势地位。本试验对野燕麦种群的繁殖与扩散特性、种子与幼苗抗逆特性及其根系分泌物对小麦的异株克生作用进行了研究,以期为找寻治理野燕麦的有效措施提供参考。结果表明:(1)野燕麦具有很强的种子繁殖与扩散能力,开花结实比小麦早,种子产量大、地下储量多、扩散距离远;(2)野燕麦种子对温度的适应范围较广,在室温与变温条件下均可以正常萌发,而冷冻后置于室温环境是其萌发最佳条件,总发芽率可达93.33%;(3)野燕麦种子对盐胁迫具有一定的适应性,在浓度低于1.8%的NaCl溶液中均可正常萌发;(4)野燕麦幼苗对盐胁迫和干旱胁迫具有较强的适应性,随着NaCl浓度和PEG浓度的升高,野燕麦幼苗叶片的脯氨酸含量显著增加(P0.01),过氧化氢酶活性呈现先增加后下降的趋势;(5)野燕麦根系水浸提液对小麦幼苗株高、干重、根长及根系活力均有极显著影响(P0.01),证明野燕麦根系分泌物对小麦幼苗生长具有异株克生作用。以上结果表明,野燕麦的竞争优势主要体现在强大的繁殖扩散能力、较强的抗逆性以及异株克生效应,相关研究可为大田野燕麦防除提供一定理论参考。     

Quantitative expression analysis of GH3, a gene induced by plant growth regulator herbicides in soybean

Symptoms resembling off-target plant growth regulator (PGR) herbicide injury are frequently found in soybean fields, but the causal agent is often difficult to identify. The expression of GH3, an auxin-regulated soybean gene, was quantified from soybean leaves injured by PGR herbicides using real-time RT-PCR. Expression of GH3 was analyzed to ascertain its suitability for use in a diagnostic assay to determine whether PGR herbicides are the cause of injury. GH3 was highly induced by dicamba within 3 days after treatment (DAT) and remained high at 7 DAT, but induction was much lower at 17 DAT. GH3 was also highly induced at 7 DAT by dicamba + diflufenzopyr, and to a lesser extent by the other PGR herbicides clopyralid and 2,4-D. The non-PGR herbicides glyphosate, imazethapyr, and fomesafen did not significantly induce GH3 expression above a low constitutive level. These results indicate that a diagnostic assay for PGR herbicide injury based on overexpression of auxin-responsive genes is feasible, and that GH3 is a potential candidate from which a diagnostic assay could be developed. However, time course analysis of GH3 expression indicates the assay would be effective for a limited time after exposure to the herbicide.     

Conserving seeds of useful wild plants in Mexico: main issues and recommendations

The efficient storage and germination of seeds underpin the effective use of plants for livelihoods and sustainable development. A total of 204 wild species useful for local communities of the Tehuacán–Cuicatlán Valley were collected and stored in seed banks in country for long term conservation, and 66 % (i.e., 134) duplicated in the U.K., as an effective means of ex situ conservation. Of the 204 species, 147 (122 of which also duplicated in the U.K.) were previously listed as useful plants in the ethnofloristic inventory of the Valley. Based on literature surveys, we found that one of the major impediments to the use of stored seeds of wild species is the lack of knowledge of how to germinate the seed. In detailed studies, we found that seeds of 18 useful plant species from 10 different families germinated readily and could be propagated. In contrast, four species (Actinocheita filicina, Bursera submoniliformis, Karwinskia mollis and Lippia graveolens) produced dormant seeds and therefore further studies are needed before their use can be maximised in large scale propagation programmes in support of conservation and livelihoods. Overall, this large-scale study on useful wild plant species in Mexico confirms that conventional seed banking can effectively support sustainable development and livelihood programmes.     

Effects of Sulfur and Water Supply on Quantitative and Qualitative Traits of Indian Mustard

Oilseed production can be affected in arid and semi-arid regions that are exposed to water shortages. Nevertheless, cultivation of drought-resistant crops such as Indian mustard is a suitable way to gain acceptable yields. Effects of different levels of irrigation and sulfur (S) fertilizer on the quantity and quality parameters of mustard (Brassica juncea) were assessed in a trial at the Indian Agricultural Research Institute, New Delhi, during 2007–2008 and 2008–2009 growing seasons. The experiment was carried out in a split plot as randomized complete block design with three replications. The experiment treatments consisted of three irrigation levels [no irrigation, one irrigation at 45 days after sowing (DAS), and two irrigations at 45 DAS and 90 DAS] in main plots and four sulfur levels (S; 0, 15, 30, and 45 kg S ha^?1) in subplots. The results showed that in the two successive years of the experiment, the number of siliquae/plant, length of siliqua, number of seeds/siliqua, and 1000-seed weight increased significantly with increasing the level of irrigation, which resulted in greater seed yield than no irrigation. Also, mustard plants irrigated two times showed the greatest values of nitrogen (N), protein, and oil content of the seeds over no irrigation in both years of this study. The oil yield and S content in the seed of mustard increased significantly with increasing the level of irrigation in both years. Number of siliquae/plant, siliqua length, number of seed/siliqua, and 1000-seed weight increased significantly with increasing the rates of S up to 45 kg S ha^?1 in both years of experimentation; however, in 2008–2009 the difference between application of 30 and 45 kg ha^?1 S was not statically significant. Also, the seed yield, N, protein, oil, and S contents in seeds of mustard increased significantly with increase in the level of applied S in both years.     

氮硫对腌制叶用芥菜营养品质的影响

通过对不同氮硫处理的两个品种叶用芥菜(雪里蕻和包包青菜)进行腌制加工,研究氮硫对其营养品质的影响。结果表明,增加氮肥量,腌制叶用芥菜的可溶性总糖、还原糖和蔗糖含量显著降低,而其可溶性蛋白质、硝酸盐和亚硝酸盐含量则显著增加;增加硫肥量,腌制叶用芥菜的蔗糖含量、可溶性蛋白质和腌制包包青菜硝酸盐含量显著增加,而叶用芥菜的亚硝酸盐含量则显著降低。两个品种相比较,腌制雪里蕻的还原糖含量、可溶性蛋白质和硝酸盐含量显著高于腌制包包青菜,相反,腌制包包青菜的亚硝酸盐含量则显著高于腌制雪里蕻。品种差异及氮、硫处理对腌制叶用芥菜中营养元素(N、S、P、K、Ca、Mg、Fe)含量均有显著影响,并呈现不同的变化规律。