Latent entry and spread of Colletotrichum acutatum (species complex) in strawberry fields

Anthracnose, caused by Colletotrichum acutatum (species complex), has become a troublesome problem in strawberry production worldwide. This paper reports (i) an optimized sampling method combined with a real‐time PCR technique to detect the latent presence of C. acutatum in cold‐stored strawberry plants used as planting material in several European countries, and (ii) a study of the spread of C. acutatum following a point inoculation under field conditions. Screening of different parts of planting material suggested that C. acutatum is most likely to be present on runners and old petioles. In addition, in seven out of nine batches of planting material from different nurseries, latent infection by C. acutatum was detected in at least one of five replicate samples. Field experiments in 2009 and 2010 showed extensive latent within‐field spread of the pathogen on strawberry leaves, with a within‐row dispersal distance up to at least 1·75 m in 1 week. A straw ground cover between the rows did not decrease C. acutatum spread, probably because introduced (and/or subsequent) inoculum was confined to the plant bed (within the row) and was not present between the beds. Moreover, the number of C. acutatum spores on the symptomless leaves, as estimated using a real‐time PCR method, was significantly (P < 0·05) correlated with the incidence of fruit rot at harvest and post‐harvest (r = 0·56–0·66). These results illustrate the importance of detecting latent infections in planting material and strawberry leaves in the field.     

Molecular cloning and expression of kidney‐type glutaminase from common carp (Cyprinus carpio) and its up‐regulation by glutamine in primary culture enterocyte

Glutaminase (GLS) is the key enzyme of glutamine (Gln) metabolism and utilization. In this study, a cDNA encoding GLS protein was identified from common carp Cyprinus carpio intestine. The open reading frame of GLS cDNA encodes a polypeptide of 595 amino acids, which shows a high similarity with its zebrafish Danio rerio counterpart. Bioinformatic analysis showed the protein belongs to kidney‐type GLS. The putative protein has glutaminase domain and ankyrin repeats domain, which are highly conserved among vertebrate orthologues. Real‐time quantitative PCR analysis revealed that the abundance of GLS mRNA was the highest in the white muscle, followed by the brain, eyeball and pituitary. Glutaminase was ubiquitously expressed in all intestinal segments of common carp. The activity of GLS did not distribute uniformly along the entire length of the intestine. In primary culture enterocyte, and the expression of GLS mRNA is up‐regulated quickly and effectively by Gln.     

Dietary methionine requirement of juvenile blunt snout bream (Megalobrama amblycephala) at a constant dietary cystine level

A 9‐week feeding trial was conducted to investigate the dietary methionine requirement of juvenile Megalobrama amblycephala at a constant dietary cystine level. Six semipurified diets were formulated to contain graded dietary methionine levels from 3.9 to 15.4 g kg^?1 in about 2.5 g kg^?1 increments in the presence of 2.2 g kg^?1 cystine. Results showed that specific growth rate (SGR) and protein efficiency ratio (PER) significantly increased with increasing dietary methionine levels from 3.9 to 12.4 g kg^?1 and thereafter kept stable. Maximum protein productive value (PPV), nitrogen retention efficiency (NRE) and liver weight were observed in 8.5 g methionine kg^?1 diet. Protein contents in whole fish body were positively correlated with dietary methionine level, while lipid contents were negatively correlated with it. Morphological index and hepatic glutamate‐pyruvate transaminase (GPT) activities were independent of dietary methionine levels. However, dietary methionine supplementation significantly improved haematological parameters, plasma methionine and total essential amino acid contents and hepatic glutamate‐oxaloacetate transaminase (GOT) activities. Analysis of dose response using broken‐line regression on the basis of SGR and PPV versus dietary methionine level estimated the optimum dietary methionine requirements of juvenile M. amblycephala to be between 8.5 and 8.4 g kg^?1 of diet (25.0 and 24.7 g kg^?1 of protein) in the presence of 2.2 g kg^?1 cystine, respectively. Hence, the corresponding total sulphur amino acids requirements of this species were calculated to be 10.7 and 10.6 g kg^?1 of diet (31.5 and 31.2 g kg^?1 of dietary protein).     

紫花苜蓿组织解剖结构对NaHCO3盐碱胁迫的响应

为研究苜蓿组织结构对NaHCO₃胁迫的响应,本实验采用石蜡切片技术和光学显微技术,以甘农3号紫花苜蓿为研究试材,比较研究了0,100和150 mmol/L的NaHCO₃胁迫处理对甘农3号紫花苜蓿根、茎和叶内部结构的影响。最显著的结果在于NaHCO₃盐碱胁迫对紫花苜蓿植株维管系统影响较大,使贯通于根、茎、叶3个器官的维管组织变小,导致木质部和韧皮部输导能力锐减,极大地限制了由根系吸收的溶解有盐离子的水分向地上部分的运输。此外,在NaHCO₃盐碱胁迫下,各营养器官的其他组织也发生了相应的改变。盐碱胁迫使苜蓿叶片整体变薄,对海绵组织的影响强于栅栏组织,造成栅栏组织在叶片中所占比例相对增加;使茎的横切面变为不规则形,表皮细胞变小变薄,胞壁角质层加强,且使位于茎中央的髓薄壁组织细胞直径减小,数量增加,而髓细胞的内含物浓度增加;在盐碱胁迫下促进了根部的发育,根部直径显著变粗,木质导管直径显著变小但数量增多。     

Genetic diversity and identification of race 3 of <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">lactucae</Emphasis> in Taiwan

Sorghum is an important drought tolerant crop cultivated for food and fodder purposes. Anthracnose disease caused by Colletotrichum graminicola is a major constraint in sorghum productivity in India. Certain antagonistic fungi, that were isolated in the previous study from the rhizosphere and rhizoplane of perennial grasses in India, were studied for their antagonism in vitro to C. graminicola, root colonization ability and rhizosphere competence. Out of 138 isolates tested, 89 were antagonistic. Fifteen fungal isolates with greater than 70 % in vitro inhibition zone to the pathogen tested positive for root and rhizosphere colonization abilities. Three isolates – Chaetomium globosum isolate 57, Trichoderma harzianum isolate 184 and Fusarium oxysporum (NSF isolate 9) with prominent biocontrol potentials were tested for the control of sorghum anthracnose in greenhouse and field. Chaetomium globosum, Trichoderma harzianum and Fusarium oxysporum isolates decreased seedling mortality, and incidence and severity of disease at different growing stages. They promoted plant growth (dry biomass- 45.3, 40.0 and 46.7 %) and increased yield (grain biomass- 33.3, 23.8 and 49.2 %) respectively, over control in field. The population of the above fungi in soil was moderately high at harvest stage. The present investigation revealed that fungal isolates from rhizosphere and rhizoplane of perennial grasses could be employed to manage anthracnose and enhance plant growth and yield potentialities in sorghum, at the same time.     

The limits of the binary power law describing spatial variability for incidence data

Taylor's power law (TPL), an empirical law relating the observed variance to mean density (or abundance), has found wide applicability for characterizing heterogeneity in many disciplines. However, when the density variable has an upper bound, the TPL does not hold and the binary power law (BPL) needs to be used instead. The BPL has been shown to describe the heterogeneity of numerous plant disease epidemic systems. In this study, a generic stochastic simulator was used to study the extent to which the BPL can satisfactorily describe incidence data. Results showed that the symmetrical BPL does hold whenever there is a positive correlation among neighbours on the probability of a plant becoming infected, or where disease development is not influenced by the neighbours. These results held for a wide range of neighbourhood sizes, strengths of neighbourhood influence, and size of the sampling quadrats. However, the symmetrical BPL did not hold when there is a negative influence among neighbours. The more general asymmetrical BPL (ABPL) fitted the data with positive or negative neighbourhood influence, but because a negative neighbourhood effect is generally unlikely for plant epidemics, the symmetrical BPL is preferred over the ABPL because of its parsimony. The magnitude of the estimated BPL parameters increased with increasing neighbourhood influence and sampling‐quadrat size. However, except when the power parameter equals 1, inferring specific underlying mechanisms generating the data or comparing BPL estimates from different studies is difficult, because of the large effect of sampling on the BPL parameter estimates.     

Epiphytic colonization of Ustilaginoidea virens on biotic and abiotic surfaces implies the widespread presence of primary inoculum for rice false smut disease

Ustilaginoidea virens (Uv), the causative agent of rice false smut disease, infects developing rice spikelets at the booting stage, and transforms individual grains of the panicle into smut balls. Epidemics of the disease occur when the rice booting and heading stages coincide with rainy days. Using a green fluorescent protein (GFP)‐labelled Uv isolate that can form false smut balls on rice panicles, it was found that under high humidity and free water conditions the Uv isolate could colonize leaves of plants belonging to various families including the Poaceae (Oryza sativa, Echinochloa crusgalli, Digitaria sanguinalis and Leptochloa chinensis), the Brassicaceae (Arabidopsis thaliana) and the Solanaceae (Nicotiana benthamiana) without symptoms. Over several days, some conidia could germinate on the leaves of these plants and in water on the surface of Parafilm and cellophane, form hyphae and differentiate conidiophores to generate a large number of secondary conidia, while other conidia were able to directly produce secondary conidia. Conversely, in the absence of water some conidia could either bud to form new conidia or were converted into chlamydospores. These data indicate that Uv is one of a few fungal pathogens reported to have epiphytic characteristics. The rapid generation of a large number of spores on biotic and abiotic surfaces greatly increases the inoculum that can infect rice spikelets, resulting in the occurrence of rice false smut disease epidemics. These findings are important in the development of disease control strategies.     

Development and identification of a introgression line with strong drought resistance at seedling stage derived from Oryza sativa L. mating with Oryza rufipogon Griff

Drought stress is severely damaging during seedling stage of rice (Oryza sativa L.), which can lead to significant yield reductions. Dongxiang common wild rice (Oryza rufipogon Griff., hereafter referred to as DXWR), with strong drought resistance, could be a favorable genetic resource to improve the drought resistance of cultivated rice. Xieqingzao B (O. sativa L. ssp. indica, hereafter referred to as XB) is a representative maintainer line in hybrid rice breeding system in China. By using DXWR as donor parent, XB as recurrent parent, through continuous selfing, backcrossing and strict drought-resistant screening, we developed a strong and stable drought-resistant introgression line IL395 (BC₅F₁₀), whose ability of drought resistance was significantly increased than that of the recurrent parent XB at the seedling stage. Meanwhile, no significant differences existed among other major agronomic traits under normal condition, except for plant height. Physiological assessment revealed that IL395 exhibited a significant increase in levels of free proline and soluble sugars, which was associated with drought resistance. Whole genome marker analyses identified genomic segments of DXWR linking with RM171 and RM590 (chr. 10) and RM235 (chr. 12) that require further analysis as possible sources of drought resistance trait. These results suggest that DXWR could be a favorable genetic resource to improve the drought resistance of cultivated rice, and the IL395 might be a useful resource for excavating the drought-resistant genes from DXWR.     

A Mediterranean japonica rice (Oryza sativa) cultivar improvement through anther culture

Certified seed producers systematically select and propagate registered varieties year after year in order to maintain their uniformity and the original registered cultivar traits. However, natural mutations, spontaneous breeding between varieties and alien grain contamination can introduce undesirable variability. NRVC 980385 is a temperate japonica rice cultivar (Oryza sativa ssp. japonica) first registered in Spain in 2002. In 2005 certification tests detected a plot differing from the original traits in terms of uniformity and height suggesting the presence of a certain heterozygosis. This material was therefore seen as an opportunity to obtain newly stabilized doubled haploid (DH) lines which could compete in the Spanish short grain seed market. In this study, an in vitro anther culture protocol is defined which also covers the field tests selection to obtain four new, improved and stabilized DH derived lines ready to be registered for commercial proposes. This took just 4 years from the initial anther collection until new lines were grown in large scale field trials. Consequently, this protocol reduces the time for obtaining field assessed DH lines thereby having considerable advantages over other techniques by both maintaining the original registered cultivars and/or generating new derived varieties.