First molecular genotyping of voltage gated sodium channel alleles in Culex quinquefasciatus populations in Malaysia

A nationwide investigation was performed to detect the presence of 1014 mutation(s) in voltage gated sodium channel (kdr) gene of Culex quinquefasciatus from 14 residential areas across 13 states and a federal territory in Malaysia. Molecular genotyping of kdr mutation was performed via a modified three tubes allele-specific-polymerase chain reaction (AS-PCR) and direct sequencing of kdr gene. Based on the results of AS-PCR, homozygous susceptible (SS) genotype was found in nine out of 14 populations with 38 individuals from a total sample size of 140. Heterozygous (RS) genotype was most predominant (99 individuals) and distributed across all study sites. Homozygous resistance (RR) genotype was detected in Perak (one individual) and Selangor (two individuals). The resistance kdr allele frequencies ranged from 0.1 to 0.55, with the highest being detected in Cx. quinquefasciatus population from Selangor. This study has documented the first field-evolved instance of 1014F mutation in Malaysian mosquitoes and the findings of this study could be utilized in the implementation of strategic measures in vector control programs in Malaysia.     

Management of roadside populations of invasive Ambrosia artemisiifolia by mowing

Ambrosia artemisiifolia (common ragweed) is a highly allergenic alien weed in Europe, which spreads rapidly along roadsides. Road verges are subject to frequent mowing, which further increases the spreading of the plants' seeds. Ambrosia artemisiifolia reacts to cutting by producing new shoots, which are able to develop flowers and ultimately new seeds. An effective mowing regime that would decrease the production of seeds and their dispersal is desirable to control the spread of the plant, but an appropriate way of mowing has yet to be found. In this study, we explored how the reproductive traits of A. artemisiifolia plants in seven spontaneous roadside populations reacted to the application of different mowing regimes over 3 years. The mowing regimes that were applied differed in the timing and frequency of cuttings. We found that the cutting regime, if appropriately timed, can strongly influence the production of male inflorescences (i.e. allergenic pollen), of female flowers (i.e. seeds) and had an impact on the phenological development of the plant. Based on our findings, we suggest that the optimal management of the plant along roadsides must be adjusted to its phenological development. The most effective mowing method of control consists of a first cut shortly before male flowering, to limit the quantities of released pollen, followed by subsequent cuts before the onset of new flowers on the resprouting lateral shoots.     

Allelopathic effect of itchgrass (Rottboellia cochinchinensis) on seed germination and plant growth

Herbicides produce a wide range of toxic side-effects that pose a potential hazard to the environment. The development of natural allelochemicals is one method of addressing these issues. Here, we describe the allelopathic activity of itchgrass ( Rottboellia cochinchinensis ), which can inhibit the seed germination and growth of weeds. Farmers in Lampang, northern Thailand, have been cultivating itchgrass and using it as a mulching material to control other weeds in vegetable fields. It has long been observed that itchgrass interferes with the growth of other plants. This study showed that the density of weed species in the itchgrass-infested areas was lower than that in the itchgrass-uninfested areas. The shoot and root growth of Bidens pilosa , Mimosa pudica , Ageratum conyzoides , Echinochloa crus-galli , Oryza sativa var. RD 6, and Lactuca sativa var. OP were significantly reduced in soil previously planted with itchgrass. Water-soluble extracts from all parts of itchgrass had inhibitory effects on the growth of some test plants. Allelochemicals from itchgrass can inhibit seed germination and plant growth better at a 1 cm distance than at a 3 cm and 5 cm distance from itchgrass. Our results suggest that itchgrass has a strong competitive ability and possible allelopathic activity to other plant species. The allelopathic activity of itchgrass in the soil can influence the germination of adjacent species, causing reduced growth of seedlings.     

Characterization of Malaysian Pectobacterium spp. from vegetables using biochemical, molecular and phylogenetic methods

In August 2011, vegetable crops showing symptoms of maceration and water soaked lesions on their tuber, leaf, and fruit were collected from four major vegetable growing states in Malaysia including Pahang, Johor, Melaka and Selangor. The majority of the causal organisms isolated from infected tissues (52 strains) were identified as Pectobacterium spp. based on PCR amplification of the pectate lyase (pel) gene and amplification of the 16S-23S rRNA (ITS) with G1 and L1 primers. Physiological and biochemical assays divided Malaysian Pectobacterium species into two main groups: Pectobacterium wasabiae and Pectobacterium carotovorum subsp carotovorum. Partial sequence of PCR product from reaction of putative Pectobacterium spp. with 16S rRNA confirmed the results obtained from physiological and biochemical assays used for identification of the bacterium. Application of specific primers such as Eca1F/Eca2r, Br1f/L1r, EXPCCF/EXPCCR, and also ITS-PCR following by RFLP by restriction enzyme (RsaI) successfully differentiated Malaysian P. wasabiae and P. carotovorum subsp carotovorum isolates from other species and subspecies of Pectobacterium. Phylogenetic analysis of Malaysian isolates with housekeeping genes (mdh, gapA) grouped Malaysian P. carotovorum subsp carotovorum and P. wasabiae in the same cluster with P. carotovorum subsp carotovorum (Ecc380) and P. wasabiae (SCRI488) respectively.     

Allelopathic potential of itchgrass (Rottboellia exaltata L. f.) powder incorporated into soil

Itchgrass ( Rottboellia exaltata L. f.) is a widespread weed in northern Thailand. The farmers in this area have been using itchgrass as a mulching material in order to control other weeds in vegetable fields. Laboratory experiments were undertaken to investigate the phytotoxic activity of itchgrass powder incorporated into soil in order to evaluate the allelopathic activity in the field. The phytotoxic activity on the growth of radish seedlings ( Raphanus sativa L. var. radicula ), used as a test plant, was more pronounced in the root than in the shoot growth. The phytotoxic activity was found to be similar for the soils incorporated with the shoot or the root powder of itchgrass. The growth of the radish seedlings grown in sea sand and watered with soil water obtained from the soil previously incorporated with itchgrass powder showed a similar inhibition to those planted in the treated soil. The phytotoxic activity on the growth of the radish seedlings in the soil incorporated with the powder decreased over time. It is suggested that itchgrass releases phytotoxic compound(s) into soil water and the concentration of the active compound(s) in the soil water decreases over time.     

The Genetic Structure of Australian Populations of Mycosphaerella musicola Suggests Restricted Gene Flow at the Continental Scale

ABSTRACT Mycosphaerella musicola causes Sigatoka disease of banana and is endemic to Australia. The population genetic structure of M. musicola in Australia was examined by applying single-copy restriction fragment length polymorphism probes to hierarchically sampled populations collected along the Australian east coast. The 363 isolates studied were from 16 plantations at 12 sites in four different regions, and comprised 11 populations. These populations displayed moderate levels of gene diversity (H = 0.142 to 0.369) and similar levels of genotypic richness and evenness. Populations were dominated by unique genotypes, but isolates sharing the same genotype (putative clones) were detected. Genotype distribution was highly localized within each population, and the majority of putative clones were detected for isolates sampled from different sporodochia in the same lesion or different lesions on a plant. Multilocus gametic disequilibrium tests provided further evidence of a degree of clonality within the populations at the plant scale. A complex pattern of population differentiation was detected for M. musicola in Australia. Populations sampled from plantations outside the two major production areas were genetically very different to all other populations. Differentiation was much lower between populations of the two major production areas, despite their geographic separation of over 1,000 km. These results suggest low gene flow at the continental scale due to limited spore dispersal and the movement of infected plant material.     

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.     

Roadside surveys support predictions of negligible naturalisation potential for cotton (Gossypium hirsutum) in north-east Australia

A survey of cotton (Gossypium hirsutum) volunteers was conducted along the 1400 km of a cottonseed transport route in north‐east Australia to determine the extent of naturalisation from spilt seed. In each year, 1% of the road distance was inspected for volunteer plants. The survey results were juxtaposed with the eco‐climatic suitability predictions from inferential modelling. Over 3 years, 22 cotton plants were found at eight sites. Within the cotton production region, volunteers averaged 2.28 plants km⁻¹ of road; their most likely source was seed cotton spilt during the previous harvest. Further north, three plants were found over 3 years, at an average density of 0.089 plants km⁻¹ of road; all three plants were found in locations with a positive Ecoclimatic Index. No secondary spread was detected. Roadside slashing reduced plant survival and the potential to produce seed. In the wet tropics, weed competition and slashing prevented volunteers establishing. The surveys indicate that roadsides in north‐east Australia are a hostile environment for the establishment of cotton plants, with no evidence of naturalisation or secondary spread. Current transport practices utilizing fully covered loads present negligible risk of producing naturalised populations of cotton (either transgenic or non‐transgenic) on roadsides in north‐east Australia.     

Invasive alien plants along roadsides in Europe

This study presents a list of invasive alien plants that are found along roadsides in seven selected European countries – Austria, Germany, the Netherlands, Ireland, Norway, Slovenia and Sweden – and an overview on the role of roadsides as a habitat for invasive alien plants. This compilation is based on national lists of invasive alien plants, a literature search and expert consultation. Comprising 89 species from 31 plant families, species introduced for horticulture dominate the list (65%). Thirteen species (14%) are widespread (occur in four or more countries) and include well‐known invasive plants such as Fallopia japonica, Heracleum mantegazzianum, Solidago canadensis and Solidago gigantea. Seventeen species are listed either on the EPPO List of Invasive Alien Plants or on the EPPO A2 List of pests recommended for regulation as quarantine pests. Five species are on the List of Invasive Alien Species of Union Concern (EU Regulation 1143/2014). The compiled list provides a snap‐shot of invasive alien plants currently found along roadsides in the selected countries. It allows for a more targeted approach to monitoring, containment and control of the most problematic invasive alien plants identified in each country. Moreover, the list may also be used to identify emerging (potentially) invasive alien plants along roadsides in other European countries that warrant monitoring and/or management.     

Phytophagous insect fauna of Onopordum spp. thistles in the south of Syria

Thistles from the genus Onopordum (Asteraceae) are of Mediterranean and Eurasian origin. They are very common in the south of Syria, in particular in Damascus and Al Suwayda, and are found in fallow fields, pastures, roadsides and neglected areas. In 2015–2017, several sites with large populations of Onopordum spp. were identified. Weekly surveys of some of the sites, mainly in the regions of Damascus and Al Suwayda, revealed a significant diversity of phytophagous insect species, some of which had a very high population density. A total of 29 insect species were detected, belonging to four orders and 21 families. The most abundant species were from the order Coleoptera. These were observed to be feeding on different plant parts of Onopordum spp. Purely endophagous species comprised 38% of the species observed on Onopordum spp., ectophagous species comprised 45% with 17% of species mixed ectophagous and endophagous. Species that only fed on Onopordum spp. represented 34% of all the species observed.     

A serious disease of seedling rambutan caused by Pseudocercospora nephelii sp. nov.

Pseudocercospora nephelii is described from necrotic lesions on leaves of Nephelium lappaceum seedlings in Brunei, sometimes causing complete defoliation and death. The pathogen also occurs in Sabah and Selangor in Malaysia. Recommendations for control include treatment with mancozeb or benomyl.     

Analysis of genetic and virulence variability of Stemphylium lycopersici associated with leaf spot of vegetable crops

Stemphylium lycopersici (Enjoji) W. Yamam was initially described from tomato and has been reported to infect different hosts worldwide. Sequence analyses of the internal transcribed spacer (ITS) regions 1 and 2, including 5.8S rDNA (ITS-5.8S rDNA) and glyceraldehyde-3-phosphate dehydrogenase (gpd) gene, random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR), as well as virulence studies were conducted to analyze 46?S. lycopersici isolates. Stemphylium lycopersici isolates used in this study were obtained from diseased tomato (Solanum lycopersicum L.), eggplant (Solanum melongena L.), pepper (Capsicum annuum L.) and lettuce (Lactuca sativa L.) from major vegetable growing regions of Malaysia, including the three states of Pahang, Johor and Selangor between 2011 and 2012. Phylogenetic analysis of a combined dataset of the ITS-5.8S rDNA and gpd regions indicated that all isolates were clustered in the sub-cluster that comprised S. lycopersici, and were distinguished from other Stemphylium species. Cluster analyses using the UPGMA method for both RAPD and ISSR markers grouped S. lycopersici isolates into three main clusters with similarity index values of 67 and 68 %. The genetic diversity data confirmed that isolates of S. lycopersici are in concordance to host plants, and not geographical origin of the isolates. All S. lycopersici isolates were pathogenic on their original host plants and showed leaf spot symptoms; however, virulence variability was observed among the isolates. In cross-inoculation assays, the representative isolates were able to cause leaf spot symptoms on eggplant, pepper, lettuce and tomato, but not on cabbage.     

Fine‐tuning of a mowing regime,a method for the management of the invasive plant,Ambrosia artemisiifolia,at different population densities

Common ragweed (Ambrosia artemisiifolia) is an invasive annual plant with highly allergenic pollen. Its spread in introduced and native ranges often occurs on roadsides, where it builds stable and rapidly growing populations. The most sustainable way of controlling the population size of this species is to prevent seed production in order to deplete the soil seed bank. Populations on roadsides are submitted to regular mowing management, which can even exacerbate the situation by inducing resprouting after cutting or by accidentally spreading seeds along the road. The population density in the juvenile stages of development could play an important role in the success of cutting regimes, as it might influence the resprouting capacity of this plant. The influence of the juvenile population density and of seven cutting regimes, differing in the timing and frequency of cuts, on easily measurable reproductive traits was investigated in a glasshouse experiment. The cutting regimes had a strong influence on the reproductive success and on the phenology of the development stages of ragweed. The population density in the juvenile stages did not play a role in further phenological development, but did influence the reproductive traits. The reproduction of ragweed can be lowered by locally adapted combinations of the timing and frequency of mowing. As the optimal management option for the reduction of both the male and female flowers, the authors suggest a first cut just before the start of male flowering, followed by subsequent cuts every 3–4 weeks.     

Classification of Italian Xanthium strumarium complex based on biological traits, electrophoretic analysis and response to maize interference

Xanthium strumarium complex was studied taking plant seeds from 16 diverse locations in Italy. Plant morphophysiological traits were noted and electrophoretical analysis of seed reserve proteins was carried out from individual plants grown at the same site, in the presence and absence of competition from maize. Significant correlations between diverse plant phenological traits, the geographical origin of the seeds and the seed protein electrophoretic patterns were found. A high correlation between seed reserve proteins and the genetic structure of this weed was also detected. Maize competition increased Xanthium stem growth and decreased bur production and number of branches. Xanthium plant populations could be classified according to the morphological characters evaluated in three groups closely related to their geographical original area and latitude (northern, central and southern Italy), each showing precise morphological traits of Xanthium italicum , X. strumarium and X. orientale respectively. Electrophoresis supported the classification of these three morphological groups.     

Phytoplasmas associated with disease of coconut in Malaysia: phylogenetic groups and host plant species

Coconut palm ( Cocos nucifera ), oil palm ( Elaeis guineensis ), Bermudagrass ( Cynodon dactylon ) and Madagascar periwinkle ( Catharanthus roseus ) with symptoms indicative of phytoplasma disease were collected from different locations in Malaysia. PCR assays employing phytoplasma universal rRNA gene primers P1/P7 alone or P1/P7 followed by R16F2n/R16R2 detected phytoplasmas in eight out of 20 Malayan Red Dwarf (MRD), nine out of 12 Malayan Yellow Dwarf (MYD) and 12 out of 12 Malayan Tall (MT) coconut palms displaying coconut yellow decline symptoms. Positive detections were also obtained from six out of six oil palm seedlings showing symptoms of yellowing and necrosis, from 10 out of 10 Bermudagrass samples with white leaf symptoms, and from eight out of eight periwinkle plants showing phyllody, virescence, little leaf, proliferation and foliar yellowing. Phytoplasmas were not detected in any of the symptomless plants tested. Sequencing and phylogenetic analysis of PCR products determined that phytoplasmas infecting both MRD and MT coconuts and Bermudagrass in Serdang, Selangor State, were all members of the 16SrXIV ' Candidatus Phytoplasma cynodontis' group, whereas isolates in periwinkle in Serdang were all members of the 16SrI ' Ca. Phytoplasma asteris' group. However, the phytoplasmas detected in MYD coconuts and oil palms from Banting, Selangor State, and in periwinkle from Putrajaya were collectively very similar (99%), but shared <97·5% similarity with 16S rDNA sequences of all other known phytoplasmas, indicating that they represent a novel taxonomic group. Thus, at least two phylogenetically distinct phytoplasmas are associated with the coconut yellow decline syndrome in Malaysia, both of which were also detected in other plant species.     

Current status of red palm weevil in Turkey

The red palm weevil Rhynchophorus ferrugineus (Olivier) is the most serious pest of palms in Turkey. Weevil infestation was first detected in Turkey in summer 2005 in parks and gardens of Mersin province, along the Mediterreanean coast of Turkey. Following the discovery of the pest, destruction of infested plant material, prophylactic insecticide chemical treatments, as well as adult weevil trapping were carried out on palm plantations. Traps containing a commercial aggregation pheromone were hung on palm trees at a high density, in order to monitor the pest infestation and reduce the weevil population by mass trapping. A significant decrease in the number of trapped beetles and destruction of infested plant material was observed in 2009 and continued in the following years in several cities in Turkey. Therefore, it has been observed that mass trapping and curative pesticide applications have played a significant role in the suppression of R. ferrugineus populations in palm plantations.     

A tetraploid Echinochloa with plagiotropic tillers: Its distribution and habitat in the northern part of the main island of Japan

An annual Echinochloa with plagiotropic tillers (semi‐erect type) was found on a paddy levee in Miyagi prefecture in Japan. This semi‐erect type had tetraploid chromosome numbers (2n = 36) and would be expected to be E. oryzicola. However, its appearance was distinct from that of E. oryzicola. To collect seed samples and clarify its distribution and habitat, exploration and collection surveys were carried out in the northern part of Honshu, the main island of Japan in 2014, 2017, and 2018. Thirty‐two populations that were assumed to be the semi‐erect type were found and their mature seeds were collected. Based on the results of chromosome counts and growth form analyses, all of them were confirmed to be the semi‐erect type. The 32 populations were distributed in the prefectures of Aomori, Iwate, Akita, Miyagi, Yamagata, Fukushima, and Niigata, and located in paddy fields, abandoned paddy fields, paddy levees, roadsides, vacant land, and scree slopes. Our surveys indicated that the semi‐erect type grew in wet and dry disturbed habitats at an early stage of secondary succession including paddy fields in the northern part of the main island of Japan.     

The role of arable weed seeds for agroecosystem functioning

A literature study was conducted to gather knowledge on the impact of weed seeds on agroecosystem functioning other than effects related to the production of weed seedlings and plants. The results of the review suggested that a larger and more diverse weed seedbank can contribute to the biodiversity of various groups of macrofauna and microbiota, with a positive or negative impact on the agroecosystem. However, relationships between weed seed availability and functional biodiversity in the field have generally not been established, with the exception of case studies of seeds sustaining populations of granivorous farmland birds or acting as reservoirs and vectors of plant pathogens. To value the contribution of different weed seeds to sustaining populations of functional biota, more detailed information on their relationships with seeds is essential. Hypotheses and related questions that can be used to explore the impact of weed seeds on functional biodiversity have been identified. The identification of weed seed species that are a key to sustaining functional biota may lead to a weed management strategy aiming to minimise the damage of weeds to crops while taking the alternative ecological roles of weed seeds into account.     

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。     

Influence of tillage, straw disposal system and seed return on the population dynamics of Alopecurus myosuroides Huds. in winter wheat

Alopecurus myosuroides Huds. was studied over a 2-year period in winter wheat established after tine cultivations or direct drilling. Straw was removed by baling or spread and burnt. Seed production was either allowed or prevented by cutting and removing all vegetation at the end of the first year. Cultivation differences had no consistent effect on plant or seed populations. Straw burning destroyed about 50% of seeds and encouraged the germination of surviving seeds. Weed populations in the crop were lower on burnt than on baled areas. Where seed shedding was allowed, populations of seeds in soil and plants increased by up to nine-fold per year. Straw burning resulted in smaller population increases. Seed decline in the soil averaged about 80% per year, so that less than 6% of the weed seeds sown were still viable after 2 years’burial in the soil. Most of the seed decline occurred between July and October and was slightly greater on burnt than on baled areas. Only part of this seed loss was accounted for by germination and emergence of seedlings during summer and autumn. Plants emerging in the crop represented less than 26% of viable seeds present in the soil at time of drilling the crop. Few seedlings emerged in spring. The viability of shed seeds varied with year and with weed density. High infestations were associated with lower seed viability and also fewer heads per plant.