DNA methylation changes in fusarium wilt resistant and sensitive chickpea genotypes (Cicer arietinum L.)

DNA methylation plays an important role in the regulation of gene expression in biotic and abiotic stresses. In the present study, a methylation-sensitive amplified polymorphism (MSAP) analysis was performed to profile DNA methylation changes in seven resistant and sensitive chickpea genotypes following inoculation with Fusarium oxysporum f. sp. ciceris. In all, 27468 DNA fragments, each representing a recognition site cleaved by either or both of two isoschizomers, were amplified using nine selective primer pairs. DNA methylation was evaluated in leaves, stems and roots in control and inoculated plants. Extensive cytosine methylation alterations were found in the pathogen-treated genotypes compared with the corresponding control, including hypermethylation and demethylation as well as the potential conversion of methylation types. For all genotypes, the percentage of demethylated sites were more than methylated sites in infected plants compared with the corresponding control. No significant differences were observed for banding patterns in infected and control leaf tissues, while the differences between percentage of unchanged, methylated and demethylated sites were significant in stem and root tissues. The total numbers of methylated polymorphic bands ranged from 137 to 154 bands in Sel95th1716 and Arman, accounting for 36.81%–44.64% of all bands, respectively. Ten fragments that were differentially amplified between infected and control plants were isolated and sequenced in three tissues separately. Most of sequenced fragments showed homology with disease related genes in GenBank. The results suggest that significant differences in cytosine methylation exist between resistant and sensitive chickpea genotypes, and that hypermethylation or hypomethylation of specific genes may be involved in the chickpea resistance to Fusarium wilt.     

Ion flux kinetics in blue light‐grown field dodder (Cuscuta campestris) seedlings

Previous studies have revealed that blue light stimulates coiling and prehaustoria development in young de‐etiolated dodder seedlings prior to host attachment. In this study, the Ca²⁺ and H⁺ flux kinetics that are associated with blue light‐mediated coiling and prehaustoria development in de‐etiolated dodder seedlings were measured uninvasively using the ion‐selective, vibrating microelectrode ion flux system. The findings showed that blue light induces rapid transient changes in the net Ca²⁺ and H⁺ fluxes. A clearly pronounced lag of about several minutes was observed between the blue light‐induced changes in the Ca²⁺ and H⁺ fluxes, with H⁺ being a leading ion. The addition of 1 mol L^?1 vanadate, a known blocker of the plasma‐membrane H⁺‐ATPase, to the bathing solution completely prevented both the H⁺ and the Ca²⁺ flux responses and inhibited coiling and prehaustoria development, suggesting a causal relationship between the blue light‐induced changes in the net Ca²⁺ and H⁺ fluxes and seedling movements. It is concluded that the plasma‐membrane H⁺‐ATPase plays a key role in a dodder's parasitic mode of growth and its adaptive response to the environment.     

The parasitoid complex ofLiriomyza cicerina on chickpea (Cicer arietinum)

Liriomyza cicerina (Diptera: Agromyzidae) is an important pest on chickpea in Turkey. The objective of this study was to determine the parasitoids and rates of parasitism ofL. cicerina on chickpea (Cicer arietinum L.) during the 2005 and 2006 seasons in ?anl?urfa province, Turkey. Leaves with mines were sampled weekly and kept in the laboratory to observe and count emerging leafminer and parasitoid adults. Eight parasitoid species were collected: the braconidsOpius monilicornis Fischer andOpius tersus Foerster and the eulophidsDiaulinopsis arenaria (Erdös) andNeochrysocharis formosa (Westwood), which occurred in both the winter and summer seasons;Diglyphus crassinervis Erdös,Neochrysocharis ambitiosa Hansson,Neochrysocharis sericea (Erdös) andPediobius metallicus (Nees), which occurred only in the summer growing areas.Diaulinopsis arenaria was the predominant parasitoid with 4–7.7% parasitism rate whileN. ambitiosa andO. monilicornis were the second and third most predominant species. The results of these trials show that sinceDia. arenaria occurred throughout every season, it could potentially be used for control of the leafminerL. cicerina.     

Pathogenic diversity within field populations of Orobanche cumana and different reactions on sunflower genotypes

Different races of the parasitic Orobanche cumana (sunflower broomrape) have been reported in Spain, race F being the most virulent. Full resistance in sunflower to races A–E is achieved with each of the single major genes Or₁ to Or₅ respectively. However, parasitised hybrids allegedly resistant to race F were observed in early 2002. The purpose of this study was to verify broomrape incidences (BI) on resistant sunflower genotypes, to assess the mixture of races within field populations and to test for partial resistance to race F in the sunflower hybrids showing a low degree of attack (DA) by the weed. Tests were conducted under field conditions in two locations of southern Spain. While no significant differences were found for yield and BI between locations, the DA on the cultivars depended on the location. With high infection levels and significantly lower yield in susceptible controls, marked differences in BI and DA were found within resistant cultivars, but all of them showed similar crop yield. When artificially inoculated with several populations of race F, line P96 and mainly line L86, were consistently slightly infected, suggesting they were inbred lines responsible for horizontal resistance in infested fields. L86 was extremely susceptible to race E populations, which is unusual as sunflower resistance to one race provided resistance to all the previously described races of O. cumana. No different virulences were detected within two groups of subpopulations (races E and F) inoculated onto resistant sunflower genotypes. However, race F subpopulations showed significant differences in aggressiveness, which seems to be related to horizontal (multigenic) resistance of the crop to the parasitic weed.     

Assessment of yield losses caused by chickpea chlorotic dwarf geminivirus in chickpea (Cicer arietinum) in India

Yield losses caused by chickpea chlorotic dwarf virus in chickpea were estimated by comparing uninfected and infected plants in the field at two locations in India. When infection was before flowering, yield losses of individual plants amounted to nearly 100% in the three cultivars studies. Plants that became infected during flowering had yield losses of 75–90%. Percentage of crop loss is likely to equal percentage of disease incidence, since plant densities in farmers' fields are probably too low to allow uninfected plants around infected ones to compensate the yield losses of infected plants.Submitted as Journal Article No. 1624 by ICRISAT.     

Influence of nitrogen fertilization and isoxaflutole on the nodulation of chickpea (Cicer arietinum)

The effects of soil nitrate and the herbicide, isoxaflutole, on chickpea nodulation were examined in a glasshouse experiment. The treatments consisted of one isoxaflutole‐tolerant and one isoxaflutole‐sensitive chickpea cultivar, five nitrate concentrations (0, 0.75, 1.5, 3.0, and 6.0 mmol L^?1), and three herbicide rates (0, 75 [the recommended rate], and 300 g ha^?1). The sensitive cultivar was more susceptible to isoxaflutole damage with an increasing herbicide rate and nitrate concentration, compared with the tolerant cultivar. Isoxaflutole at the recommended rate reduced the nodule dry weight of the sensitive cultivar by 51% and by 33% in the tolerant cultivar. The nodule dry weight of the sensitive cultivar was reduced by 28%, 40%, 64%, and 76% with the addition of the 0.75, 1.5, 3.0, and 6.0 mmol L^?1 nitrate concentrations, respectively. In comparison, the nodule dry weight of the tolerant cultivar was reduced by 50% and 79% at the 3.0 and 6.0 mmol L^?1 nitrate concentrations, respectively. In addition, the level of damage from increasing rates of isoxaflutole increased with the highest nitrate concentrations of 3.0 and 6.0 mmol L^?1, regardless of the cultivar. Isoxaflutole at the recommended application rate had a detrimental effect on the nodulation of both chickpea cultivars, but the isoxaflutole‐tolerant cultivar suffered less injury than the sensitive one in relation to some growth parameters. The damage to chickpea from the recommended rate of isoxaflutole also increased with soils of higher nitrate content.     

Influence of obligate parasite Cuscuta campestris on the community of its host Mikania micrantha

As a means of biologically controlling Mikania micrantha in South China, the influence of the native obligate parasite Cuscuta campestris on its natural community was studied in the field. Mikania micrantha is a non‐indigenous vine that smothers other vegetation and has become a major invader of agricultural land and native areas in Southern China. These preliminary results showed pronounced effects on M. micrantha by C. campestris. Cuscuta campestris significantly reduced biomass of M. micrantha, increased species diversity and helped re‐establishment of native species. Biomass of M. micrantha decreased from 328 g m^?2 to 82 g m^?2, biomass of companion species increased from 41 g m^?2 to 145 g m^?2, the total number of species increased from 7 to 19 and the species diversity index from 1.8 to 5.6, when C. campestris was present. These results indicated that the use of C. campestris could be a potentially effective way of controlling M. micrantha and could help us achieve the novel objective of biological control of weeds using weeds.     

Allelopathic potential of wild onion (Asphodelus tenuifolius) on the germination and seedling growth of chickpea (Cicer arietinum)

Water extracts obtained from the roots, shoots, and fruits of mature wild onion ( Asphodelus tenuifolius ) plants and soil taken from an A. tenuifolius field were used to determine their allelopathic effects on the germination and seedling growth of chickpea ( Cicer arietinum ) in the laboratory. The roots, shoots, and fruits of A. tenuifolius were soaked individually in water in a ratio of 1:20 (w/v) for 24 h to prepare the extracts. Distilled water was used as the control. The germinated seeds were taken out from the Petri dishes and counted every day for 12 days. The seeds of chickpea were also sown in sand and in each of the controlled, normal soil and the soil taken from the A. tenuifolius -infested field in Petri dishes to record the length and weight of the roots and shoots 18 days after sowing. The mean germination time reached the maximum amount for the stem and fruit extracts. The fruit extract caused the most reduction in the germination index and the germination percentage of chickpea. The different wild onion organ extracts significantly reduced the root and shoot length and biomass of the chickpea seedlings compared with the distilled water. The fruit extract of wild onion proved to be the most detrimental to the root length, shoot length, and dry weight of the chickpea seedlings. The soil beneath the A. tenuifolius plants significantly reduced the emergence, root length, shoot length, shoot dry weight, and seedling dry weight but increased the root dry weight of the chickpea seedlings. It is suggested that A. tenuifolius releases phytotoxic compound(s).     

Herbicidal activity on acetolactate synthase‐resistant barnyardgrass (Echinochloa crus‐galli) in Arkansas,USA

The intensive use of the acetolactate synthase (ALS)‐inhibiting herbicides, imazethapyr, penoxsulam and bispyribac‐sodium, in imidazolinone‐resistant (Clearfield) rice increases the risk of the evolution of ALS‐resistant barnyardgrass. In 2009, imazethapyr failed to control barnyardgrass that was collected from a field in Arkansas, USA, following the failure of the herbicide in 2008. A greenhouse experiment was conducted to confirm and document the level of resistance of the biotype against three ALS‐inhibiting herbicides that currently are labeled in rice. The level of control of the resistant biotype at the labeled rate of bispyribac‐sodium of 35 g ai ha^?1 was 10%, penoxsulam at 22 g ai ha^?1 was 0% and imazethapyr at 70 g ai ha^?1 was 25%. The level of mortality of the susceptible biotype was 100% with all the herbicides at the labeled rate. The dose needed to kill 50% of the resistant plants was 49 g ha^?1 of bispyribac‐sodium, 254 g ha^?1 of penoxsulam and 170 g ha^?1 of imazethapyr. For the susceptible biotype, bispyribac‐sodium at 6 g ha^?1, penoxsulam at 10 g ha^?1 and imazethapyr at 12 g ha^?1 killed 50% of the treated plants. Based on these findings, it was confirmed that a barnyardgrass population has evolved cross‐resistance to three ALS‐inhibiting herbicides in rice culture in Arkansas. Furthermore, an experiment was conducted to determine if the ALS‐resistant biotype could be controlled using other mechanisms of action. The results indicated that propanil, a photosystem II inhibitor, and quinclorac, a synthetic auxin, failed to control the resistant biotype at the labeled rates, whereas all the other evaluated herbicides provided effective control of both biotypes.     

新疆鹰嘴豆不同种植密度对其生长发育产量和品质的影响

以阿瓦提1号为材料,研究新疆地区不同种植密度对鹰嘴豆植株性状、干物质积累量、产量与品质的影响。结果表明,随密度的增加,生育期提前,株高增高,叶面积指数上升,单株干物质积累量减少。单株荚数随着密度增大极显著减少。每荚粒数、粒重等与密度的关系不大。蛋白质含量、亚麻酸含量和廿碳烯酸含量都呈现增加的趋势。种植密度对鹰嘴豆维生素B1和油酸含量的影响不显著。在本试验条件下,以44.5万株/hm2处理的表现最佳,产量达5946.09 kg/hm2,品质较好。     

The effect of temperature on glutathione S-transferase activity and glutathione content in Alopecurus myosuroides (black grass) biotypes susceptible and resistant to herbicides

Herbicide‐resistant populations of Alopecurus myosuroides (black grass) have become widespread throughout the UK since the early 1980s. Previous observations in this laboratory have demonstrated that natural climatic fluctuations caused increases in endogenous glutathione S‐transferase (GST) enzyme activity in A. myosuroides plants as they mature, which is thought to be linked to herbicide resistance in this species. The present study has investigated the effects of plant growth at 10°C and 25°C, and reports GST specific activity and glutathione (GSH) pool size in resistant and susceptible A. myosuroides biotypes. Findings demonstrate differences in GST activity between resistant and susceptible populations, which are transient at lower growth temperatures. The GSH pool size was elevated at lower growth temperature in both biotypes. We speculate that these endogenous responses are part of a natural mechanism of acclimation to environmental change in this species and suggest that resistant plants are more able to adapt to environmental stress, as indicated in this instance by temperature change. These observations imply that the control of resistant A. myosuroides by graminicides may be more effective when applied at lower temperatures and at earlier growth stages.     

Root exudates associated with the resistance of four chickpea cultivars (Cicer arietinum) to two races of Fusarium oxysporum f.sp. ciceri

The germination of race 1 spores of Fusarium oxysporum f. sp. ciceri was significantly inhibited by the root exudate of the wilt-resistant chickpea cvs CPS1 and WR315 compared to untreated spores and spores treated with root exudates from susceptible cultivars. The effect was concentration dependent, such that the exudate from 1 g of root in 2 ml of water almost completely inhibited spore germination, whereas the exudate from 1 g of root in 20 ml water did not do so. The inhibitory effects of the active exudates were negated when the apolar components of the exudates were removed by extraction with ethyl acetate. The root exudates of the susceptible cv. JG62 and the late wilting cv. H208 did not inhibit germination. The hyphal growth of germinated spores was also strongly inhibited by the concentrated exudates of CPS1 and WR315, and diluted exudates were less potent. The highest concentration of the exudate of the susceptible cv. JG62 showed some inhibition of hyphal growth, whereas none of the exudates of H208 were found to contain any antifungal activity. The effect of the exudates on the spores of race 2 was similar to that reported for race 1, except that the water-soluble components of the crude root exudate of WR315 after ethyl acetate extraction were also found to inhibit germination significantly. Overall, the spores of race 2 appeared to be more susceptible to the effects of the exudates. The ethyl acetate fractions of the root exudates of CPS1 and WR315 strongly inhibited germination and hyphal growth of both race 1 and race 2, the effect being concentration dependent. The results suggest that the resistance of chickpeas to vascular wilt depends, at least in part, upon the antifungal activity of the root exudates. Differences in the expression of resistance in the field could depend upon the concentration or rate of production of constitutive antifungal components by the root.     

Preliminary findings of potentially resistant goosegrass (Eleusine indica) to glufosinate‐ammonium in Malaysia

Goosegrass (Eleusine indica), regarded as one of the world's worst weeds, is highly pernicious to cash crop‐growers in Malaysia. Following reports in 2009 that glufosinate‐ammonium failed to adequately control goosegrass populations in Kesang, Malacca, and Jerantut, Pahang, Malaysia, on‐site field trials were conducted to assess the efficacy of glufosinate‐ammonium towards goosegrass in both places. Preliminary screenings with glufosinate‐ammonium at the recommended rate of 495 g ai ha^?1 provided 82% control of the weed at a vegetable farm in Kesang, while the same rate failed to control goosegrass at an oil palm nursery in Jerantut. The ensuing greenhouse evaluations indicated that the “Kesang” biotype exhibited twofold resistance, while the “Jerantut” biotype exhibited eightfold resistance towards glufosinate‐ammonium, compared to susceptible goosegrass populations. The occurrence of glufosinate resistance in goosegrass calls for more integrated management of the weed to prevent escalating resistance and further proliferation of the weed in Malaysia.     

Solarization for the control of Pueraria montana (kudzu)

The effects of translucent polyethylene sheeting as a thermal covering to eradicate Pueraria montana (kudzu) were investigated at Clemson, South Carolina on a clay loam. In 2005, the highest reduction of live root crowns was observed where P. montana was covered for the entire growing season (May–October) with a reduction of 42% of live root crowns compared with the control plots. Where P. montana was covered for alternate weeks throughout the growing season live root crowns were reduced by 35%. Covering P. montana for one week and then uncovering for four weeks repeatedly through the season killed 24% of root crowns. In 2006, the second year of treatment to the same plots, the three solarization treatments had similar efficacy of about 97%. The use of polyethylene sheeting appears not to be cost‐effective for general control of large P. montana infestations, but may be useful for small patches.     

Identification of sources of resistance to crenate broomrape (Orobanche crenata) in Spanish lentil (Lens culinaris) germplasm

The weedy root parasite Orobanche crenata constitutes a serious threat to lentils and other legumes grown in the Mediterranean and western Asia. Control strategies have centred on agronomic practices and the use of herbicides. Resistance breeding is hampered by scarcity of proper sources of resistance and of a reliable and practical screening procedure. A stepwise procedure was used here. A Spanish germplasm collection of 234 accessions of lentils (Lens culinaris) was screened for resistance to crenate broomrape under field conditions. A wide range of responses was observed, however, complete resistance was not detected. Some accessions exhibited a substantial reduction in broomrape infection. Thirty‐five lentil accessions with reduced broomrape infection were selected for further screening in the field and to determine the components of resistance in pot and in mini‐rhizotron experiments. Low infection seemed to be based on a combination of various escape and resistance mechanisms: (i) a lower root density, (ii) a lower induction of Orobanche seed germination, (iii) the reduced establishment of broomrape radicles and (iv) a limited development of established tubercles. In addition, necrosis of tubercles was observed in some accessions. This study shows how complementary in vitro screening methods may be used to rank and identify lentil accessions with potential broomrape resistance, based on different resistance mechanisms which may in the future be combined into a single cultivar by breeding.     

Weed seedling herbivory by field cricket Teleogryllus emma (Orthoptera: Gryllidae) in relation to the depth of seedling emergence

This study aimed to determine whether seedling herbivory by the field cricket, Teleogryllus emma (Ohmachi et Matsuura), promoted the mortality of Italian ryegrass (Lolium multiflorum Lam.), a non‐native grass weed in Japan, whether the impact of seedling herbivory differed depending on the depth of seedling emergence and the influence of cricket density on seedling mortality. First, the seedlings at emergence depths of 0, 1 and 2 cm were exposed to the cricket for 2 days in plastic cups and the amount of seedling mortality by herbivory was calculated. The level of mortality of 60 seedlings that emerged from seeds on the soil surface was 92.4%, significantly higher than that of the seedlings that emerged from seeds at depths of 1 cm (10.8%) and 2 cm (9.6%). Second, the seedlings at the emergence depths of 0 and 1 cm were exposed to the crickets at four different densities (zero, one, two and three individuals per 5700 cm²) for 5 days in plastic containers and the amount of seedling mortality by herbivory was calculated. The level of mortality by herbivory of the seedlings that emerged from 100 seeds on the soil surface was higher (33.3–61.3%) than that of the seedlings that emerged from seeds at a depth of 1 cm (4.7–13.1%) in the containers with one‐to‐three crickets. The level of seedling mortality tended to increase with an increasing cricket density. These results suggest that seedling herbivory by crickets is an important factor in weed mortality, particularly in no‐till fields and in field margins, where the seeds are more prevalent near the soil surface.