Increasing maturity reduces wound response and lignification processes against Penicillium expansum (pathogen) and Penicillium digitatum (non-host pathogen) infection in apples

Penicillium expansum is the main postharvest pathogen of pome fruit and is a necrotrophic fungus that requires wounds to infect the fruit. Therefore, injuries caused during harvest and postharvest handling provide an optimal locus for infection. In this study, the effect of wound response in apples harvested at three different maturity stages and stored at two different temperatures (20 and 0 °C) infected with P. expansum (pathogen) and Penicillium digitatum (non-host pathogen) was evaluated. The effect of wounding and pathogen inoculation on lignin content was also quantified. At 20 °C, less decay incidence and severity were observed when time between wounding and inoculation increased, and these differences were more important in fruit from immature and commercial harvests. However, at 0 °C, wound response was too slow to prevent P. expansum infection. Lignin content was highest in fruit from the immature harvest. Our results indicated that maturity and storage temperature play an important role in apple wound response. This is the first report demonstrating that P. digitatum, a non-host pathogen, was able to develop rots in over-mature apples.     

Rhodosporidium paludigenum induces resistance and defense-related responses against Penicillium digitatum in citrus fruit

Induced disease resistance against plant pathogens is a promising non-fungicidal decay control strategy. In this study, a potential biocontrol yeast, Rhodosporidium paludigenum, was investigated for its induction of disease resistance against Penicillium digitatum in citrus fruit. The results showed that R. paludigenum is the most effective yeast among three selected yeasts in stimulating the resistance of citrus fruit to green mold. When R. paludigenum was applied 48–72 h before inoculation with P. digitatum, disease incidence and disease severity in citrus fruit significantly decreased. Application of R. paludigenum at concentrations of 1 × 10⁸ and 1 × 10⁹ cells mL⁻¹ respectively resulted in 49.6% and 52.5% reductions in the percentage of infections. Induction of resistance to P. digitatum by R. paludigenum treatment significantly enhanced the activities of defense-related enzymes, including β-1,3-glucanase, phenylalanine ammonia-lyase, peroxidase, and polyphenoloxidase, which may be an important mechanism by which the biocontrol yeast reduces the fungal disease of citrus fruit caused by P. digitatum.     

Rhodosporidium paludigenum induced resistance in Ponkan mandarin against Penicillium digitatum requires ethylene-dependent signaling pathway

This work determined if the ethylene dependent signal pathway was required for antagonist-mediated fruit defense mechanisms through investigation of disease resistance against Penicillium digitatum in Ponkan mandarin induced by 1-methylcyclopropene (1-MCP), an inhibitor of ethylene perception, and Rhodosporidium paludigenum. Blocking ethylene perception with 1-MCP resulted in an increase in ACS1, ACS2 and ACO expression, and consequently an increase in ethylene production during mechanical wounding and resistance induction. The expression of the ethylene receptors ETR1, ETR2 and ETR4 as well as ethylene response factor (ERF) were observed with similar responses to yeast and 1-MCP stimuli, with ETR3 mRNA accumulation being the most sensitive to yeast application while ERS1 was the least sensitive. When applied at concentrations greater than 500 nL L⁻¹, 1-MCP pre-fumigation significantly reduced the fruit's natural protection and R. paludigenum induced disease resistance to Penicillium decay, indicating that ethylene perception was involved in inducting disease resistance. Moreover, expression of the defensive genes CHI, β-1,3-glucanase, PAL and CIN up-regulated by yeast was inhibited to different degrees by the 1-MCP pre-treatment. This study provides evidence that the biocontrol yeast R. paludigenum increased disease resistance in Ponkan mandarin against P. digitatum infection due to ethylene and signaling pathway dependent mechanisms.     

Use of citral incorporated in postharvest wax of citrus fruit as a botanical fungicide against Penicillium digitatum

The antifungal activity of citral against Penicillium digitatum, the causal agent of citrus green mold, was tested by in vitro and in vivo experiments. In vitro assays showed that the minimum inhibitory concentration and the minimum fungicidal concentration (MFC) were both 4000 μL L⁻¹. Results of in vivo tests demonstrated that wax + citral (1× MFC) treatment did not effectively inhibit the growth of P. digitatum in Ponkan mandarin fruit, whereas wax + citral (10× MFC) treatment significantly decreased the incidence of green mold after 6 days of storage at 25 ± 2 °C. Wax + citral (10× MFC) treatment remarkably increased the content of vitamin C and antioxidant enzyme activities such as catalase, superoxidase dismutase, and peroxidase but decreased the activities of phenylalanine ammonia lyase, polyphenol oxidase, and malonaldehyde. The treatment had minor effects on the pH, coloration index, and total soluble solids. This study provided theoretical data for the practical application of citral on citrus fruit quality during postharvest storage.     

柑橘绿霉病菌拮抗放线菌MY-5的筛选与鉴定

从广东省各地区水果园和蔬菜地采集土壤23份,采用稀释平板法分离获得放线菌212株。以柑橘绿霉病菌为目标菌,采用抑菌带测定法,筛选获得9株具有拮抗活性的菌株,其中菌株MY-5拮抗活性最强,抑菌带宽达18.67mm,同时对其它13种果蔬采后病害也有较强的拮抗作用。通过对菌株MY-5形态观察、培养特征、生理生化反应和16SrDNA鉴定,该菌株为链霉菌属吸水类群（Streptomyces hygroscopicus）。     

Molecular insights into fungicide resistance in sensitive and resistant Penicillium digitatum strains infecting citrus

The continuous use of chemical fungicides on citrus postharvest has led to the development of resistant strains against the fungicides in use, representing a considerable threat because the control systems are no longer effective. Evaluation of the sensitivity of 75 Penicillium digitatum strains to seven different fungicides revealed the presence of a significant number of TBZ- (84%) and IMZ-resistant (77%) strains, i.e., those fungicides most used in citrus postharvest. Molecular characterization of different P. digitatum genes involved in fungicide resistance was carried out. All P. digitatum genes were selected based on particular mechanisms of resistance due to fungicide target or mode of action. TBZ-resistance was characterized by a unique point mutation in the β-tubulin gene sequence corresponding to amino acid 200, confirming previous work on this subject. Moderate to low resistance to strobilurins did not reveal any mutation in the cytochrome b gene. DMI-resistance was evaluated by examining the CYP51 gene and four different ABC transporters PMR1, PMR3, PMR4 and PMR5. The CYP51 gene did not exhibit any mutation relating to DMI-resistance, but a five tandem repeat sequence previously described was found in the CYP51 promoter in 3 of the 75 isolates examined, whereas DMI-sensitive isolates and the other DMI-resistant isolates of P. digitatum had only one tandem repeat. Of all the ABC transporters studied, only PMR1 and PMR5 appear to be involved in fungicide resistance and several mutations were found in the promoter and the coding region for PMR5 in resistant strains compared to sensitive ones. In all cases, the resistance mechanism was consistent in both orchard or packing-house isolates and no differences conferred by either origin or fungicide pressure were observed.Consequently, since different processes have been described that confer fungicide resistance to the same compounds, such as DMIs, the hypothesis that multiple mechanisms could be acting simultaneously gains strength.     

Characterization of resistance response of pea (Pisum spp.) against rust (Uromyces pisi)

Components of resistance to the pea rust (Uromyces pisi) were studied at the histological level in seedlings of seven pea (Pisum spp.) accessions showing partial resistance under field conditions. Resistance was characterized macroscopically by an increased latent period and a decreased infection frequency in spite of a compatible interaction (high infection type). Histological investigations revealed little differences among accessions in spore germination and appressoria formation, differences being more evident once the stomata were penetrated by the infection structures. Resistance was mainly due to a restriction of haustorium formation with significant levels of early aborted colonies in accessions IFPI3260, PI347321 and PI347347. Furthermore, a reduction in the number of haustoria per colony, hyphal tips per colony and smaller colony size were found in all genotypes studied. Resistance was not associated with early‐ or late‐acting hypersensitivity in any of the genotypes studied.     

The inheritance of frost resistance and flowering response in broccoli (Brassica oleracea var. Italica)

Summary Broccoli inbred lines derived from the cross, broccoli x (cabbage x kale) were observed to differ in flowering response and frost resistance. Analysis of the F₂ population from the cross, slow-bolter, frost tolerant and bolter, frost susceptible suggested that two dominant epistatic genes conditioned frost resistance, whereas flowering was polygenically inherited.Published with the approval of the Director of the Michigan Agricultural Experiment Station as Journal Article Number 4685.     

Differential response and genes for resistance to Peronospora parasitica (downy mildew) in Brassica juncea (mustard)

The response of a wide range of Brassica juncea accessions to 14 isolates of Peronospora parasitica, 12 from India (IP00A, IP02, IP03, IP04, IP04A, IP05, IP05B, IP33 and IP33A were derived from B. juncea; IP09, IP14 and IP13A from B. rapa) and two from B. napus in the UK (R1 and P003), was screened. Sixteen differential host response groups to these isolates (classified as groups A‐P) were identified. Groups‘A’and‘B’expressed the widest resistance profiles to these isolates. Group‘A’was susceptible to isolates IP05 and IP05B, moderately resistant to isolate IP33 and resistant to all other isolates. Group‘B’was susceptible to isolates IP03, IP04 and IP04A, and resistant to the other isolates. Putative homozygous lines resistant to all 14 isolates were selected from the F₄ progeny of crosses involving lines RESBJ‐200 from group‘A’(selection from cv. Kranti) and RESBJ‐190 from group‘B’(selection from cv. Krishna). Both selections were selfed and tested for uniformity of reactions to all isolates for three generations. The resistance of RESBJ‐200 to isolates IP00A, IP04A and IP33A seems to be conditioned by single dominant genes. The resistance of RESBJ‐190 to isolates IP00A, IP05B and IP33A was also conditioned by single dominant genes. The gene for resistance to IP00A and IP33A in RESBJ‐200 seems to be independent of the genes for resistance to the same isolates in RESBJ‐190. The new genes for differential resistance to P. parasitica will be of value in future studies of the genetics of the host‐pathogen interaction and for breeding for disease resistance.     

Inheritance of resistance to root-lesion nematodes (Pratylenchus thornei and P. neglectus) in five doubled-haploid populations of wheat

Nematode species Pratylenchus thornei and P. neglectus are the two most important root-lesion nematodes affecting wheat (Triticum aestivum L.) and other grain crops in Australia. For practical plant breeding, it will be valuable to know the mode of inheritance of resistance and whether the same set of genes confer resistance to both species. We evaluated reactions to P. thornei and P. neglectus of glasshouse-inoculated plants of five doubled-haploid populations derived from five resistant synthetic hexpaloid wheat lines, each crossed to the susceptible Australian wheat cultivar Janz. For each cross we determined genetic variance, heritability and minimum number of effective resistance genes for each nematode species. Distributions of nematode numbers for both species were continuous for all doubled-haploid populations. Heritabilities were high and the resistances were controlled by 4?C7 genes. There was no genetic correlation between resistance to P. thornei and to P. neglectus in four of the populations and a significant but low correlation in one. Therefore, resistances to P. thornei and to P. neglectus are probably inherited quantitatively and independently in four of these synthetic hexaploid wheat populations, with the possibility of at least one genetic factor contributing to resistance to both species in one of the populations. Parents with the greatest level of resistance will be the best to use as donor parents to adapted cultivars, and selection of resistance to both species in early generations will be optimal to carry resistance through successive cycles of inbreeding to produce resistant cultivars for release.     

一株拮抗香蕉枯萎病的青霉菌的分离及鉴定

从海南土壤中分离到一株对香蕉枯萎病病原菌有较强拮抗作用的霉菌菌株QW1,对该菌株进行了显微特征、菌落特征以及26S rDNA序列分析。该菌株在土豆葡萄糖培养基上生长的内生菌丝无横隔膜;孢子梗顶端膨大成帚状;单个分生孢子为椭圆形,蓝色;产有性孢子。菌落生长开始为白色,培养成熟后,气丝逐渐因产生孢子由白色变蓝色。以26S rDNA序列为基础构建了相关种属在内的系统发育树,其与青霉菌株的26S rDNA序列的同源性大于为99%。将该菌株鉴定为青霉菌。将该菌株和香蕉枯萎病病菌在土壤中进行了共培养研究,结果表明,该菌株在土壤中有较强的生长力,并表现出较好的对香蕉枯萎病的抗性。     

Development of a scoring scale for powdery mildew (Golovinomyces cichoracearum (DC.) V.P. Heluta) disease and identification of resistance sources in cultivated and wild sunflowers

Powdery mildew incited by Golovinomyces cichoracearum has become a serious problem on sunflower in India during the past 2–3 years. Genetic resistance in the released cultivars and the parental lines of hybrids is rather limited. Hence, screening of about 420 accessions comprising of wild Helianthus species, interspecific derivatives, core germplasm, inbred lines and few exotic accessions was done under natural field conditions for 2 years and further confirmed by screening under artificial inoculation conditions. PCR analysis using primers specific to powdery mildew causing genera gave a 391 bp band which confirmed the pathogen as G. cichoracearum. Seven different screening methods were tested which induced infection, but dusting of spores on to the healthy leaves proved to be convenient and more effective method of infection. Based on the differential response of the accessions derived from diverse genetic backgrounds, a scale for obtaining reliable estimates of the disease has been devised. Among different cultivar germplasm accessions, the disease severity index (DSI) ranged from 15 to 100 and area under disease progression curve (AUDPC) ranged from 95 to 648. Among the four groups of cultivated sunflower accessions tested, DSI and AUDPC was in the order of exotic lines < interspecific derivatives < inbred lines < core germplasm. Reliable sources of resistance to the pathogen were identified in four annual wild species (H. argophyllus, H. agrestis, H. debilis, H. praecox), six perennials (H. angustifolius, H. atrorubens, H. rigidus, H. salicifolius, H. pauciflorus and H. resinosus), two interspecific derivatives (HIR-1734-2, RES-834-3) and two exotic lines (PI 642072, EC-537925).     

The rice cultivar Baixiangzhan harbours a recessive gene xa42(t) determining resistance against Xanthomonas oryzae pv. oryzae

Bacterial leaf blight (BB) is among the “top 3” diseases in rice production. Breeding resistant cultivars has been the most effective strategy for BB management. The inbred cultivar Baixiangzhan (BXZ) showed a broad spectrum of resistance to five Xoo pathotypes in China, including the prevalent and highly virulent Xoo pathotypes Chinese Race V (CV), which can overcome the resistance of Xa4 and Xa21. The resistance heredity of BXZ has been explored in this study. A single recessive major resistance gene, which designated as xa42(t), confers resistance against the tested Xoo pathotypes CV. Linkage analysis lands xa42(t) on chromosome 6, and genetic mapping confines it to 3.9 cM region flanked by RM20558/RM20547 and RM20580. A further seven markers were developed from this interval for high‐resolution mapping, and the xa42(t) locus was narrowed to 34.8 Kb segment bounded by RM20572 and DT46. The only functionally predicted gene included in the target region is LOC_Os06g45960 if based on the ‘Nipponbare’ reference sequence. This candidate gene is predicted to encode a cytochrome P450 protein.     

Genetic basis of resistance against two Tospovirus species in tomato (Lycopersicon esculentum)

Summary A study was conducted to determine if the chlorsulfuron tolerance existing in barley could be improved with the use of in vitro tissue culture. Differential herbicide concentration led to variable culture response for callus culture.Plants regenerated from callus culture were evaluated for chlorsulfuron tolerance in the greenhouse. Regenerants showed improved tolerance when compared to the control or unselected tissue culture-derived lines.     

Somatic hybrids between navel orange (Citrus sinensis) and grapefruit (C. paradisi) for seedless triploid breeding

Protoplasts from cell suspension cultures of ‘Bonnaza’ navel orange (Citrus sinensis (L.) Osbeck) were electrically fused with mesophyll protoplasts isolated from seedless ‘Red Blush’ grapefruit (Citrusparadisi). After 6 months of culture, a total of 20 plants were regenerated. Root tip chromosome counting revealed that 4 of them were tetraploids (2n = 4x = 36)and the rest were diploids (2n = 2x = 18) morphologically resembling the mesophyll parent. After 6 months of transplantation into the greenhouse, 4 of the diploidmesophyll regenerants unexpectedly flowered, but this phenomenon disappeared in the next year. This is the first report of precocious flowering in citrus via protoplast fusion. RAPD analysis further confirmed that the tetraploid regenerants were somatic hybrids while the diploid regenerants were mesophyll parent type. This somatic hybrid will be utilized as a possible pollen parent for improving the seedy pummelo cultivars in China by producing triploid seedless pummelo hybrid. The mechanism of early flowering was also discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Andean beans (Phaseolus vulgaris L.) with resistance to the angular leaf spot pathogen (Phaeoisariopsis griseola) in southern and eastern Africa

Common beans (Phaseolus vulgaris) are separated into two distinct groups: Andean and Middle American. We identified CAL 143 as the first Andean bean with resistance to angular leaf spot disease caused by Phaeoisariopsis griseola. Angular leaf spot is the most widespread and economically important bean disease in southern and eastern Africa, and it is especially severe on the extensively grown Andean beans. Cal 143 was resistant in Malawi, South Africa, Tanzania, and Zambia, but it was susceptible in Uganda. This was attributed to the presence of races of P. griseola in Uganda not present in the other countries. We identified two additional Andean bean lines, AND 277 and AND 279, with resistance to angular leaf spot in Malawi. We also characterized the virulence diversity of 15 isolates of P. griseola from southern and eastern Africa into nine different races. Five of six isolates from Malawi and two of seven from Uganda, obtained from large-seeded Andean beans, were characterized into four different races considered Andean. These were compatible only or mostly with large-seeded Andean cultivars. The other eight isolates from Uganda, Malawi, and the Democratic Republic of Congo, obtained from a small- or medium-seeded Middle American beans, were characterized into five different Middle American races. These were compatible with Middle American and Andean cultivars. CAL 143 was resistant or intermediate under greenhouse conditions to all but one of the same 15 isolates from southern and eastern Africa, but it was susceptible to an isolate from Uganda obtained from a medium-seeded Middle American bean. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic variation and response to selection for resistance to root-knot nematodes in oil radish (Raphanus sativus ssp. oleiferus)

Based on a preliminary screening of a collection of cruciferous crops, such as oilseed rape (Brassica napus L. ssp. oleifera Metzg.), oil radish (Raphanus sativus L. ssp. oleiferus DC.), and yellow mustard (Sinapis alba L.), some plants within the oil radish varieties showed resistance to the root-knot nematodes Meloidogyne hapla Chitwood and M. incognita (Kofoid & White) Chitwood. Resistance tests of S₁ progeny of these selected genotypes revealed a quantitative nature of resistance to M. hapla. Only a few resistant individuals were found, but a significant effect of selection was observed. In contrast, the progenies of plants resistant to M. incognita showed a very low number of egg masses, suggesting that this resistance may be conferred by dominant major gene(s). The results indicate that resistance of oil radish to root-knot nematodes may be effective and may thus provide new possibilities for the management of M. hapla and M. incognita.     

Potential of Nicotiana glauca (Grah.) as a source of resistance to black root rot Thielaviopsis basicola (Berk. and Broome) Ferr. in tobacco improvement

Nicotiana glauca, a wild relative of Nicotiana tabacum, is an attractive potential source of black root rot (Thielaviopsis basicola)‐resistant germplasm. Moreover, it shows a resistance or tolerance to PVY, TEV, anthracnose and powdery mildew. In this study its potential as a source of resistance to black root rot caused by Th. basicola was investigated. Nicotiana glauca GG (2n = 2x = 24) was crossed as male with two N. tabacum TT (2n = 4x = 48) flue‐cured cultivars: ‘BY103’ and ‘K 326’, both of which are susceptible to black root rot. Amphihaploid F₁ TG (2n = 3x = 36), amphidiploid TTGG (2n = 6x = 72) and sesquidiploid TTG (2n = 5x = 60) hybrids were obtained. The resultant sesquidiploid hybrids were used as maternal components in backcrossing to N. tabacum and a segregating post‐sesquidiploid TTg (2n = 5x = 52–54) offspring was obtained. Amphihaploids exhibited a level of resistance to black root rot characteristic of N. glauca. The expression of resistance varied in the sesquidiploid generation, possibly reflecting cytological instabilities in that progeny. A wide variation in response to black root rot was found for post‐sesquidiploids a clear reflection of extensive chromosome segregation in that hybrid generation.