Culture storage age and fungal re‐isolation from host‐tissue influence Colletotrichum spp. virulence to pepper fruits

Using resistant cultivars is the most sustainable and practical approach against plant diseases. Plant germplasm and breeding lines are selected and assayed against, usually, the most aggressive or virulent strains of a pathogen (e.g., fungus) that causes the disease. However, prolong storage of the pathogen in culture media could affect virulence that, consequently, also influence the outcome of the resistance assay. This study demonstrates that long‐term storage (at least a year) of Colletotrichum truncatum and C. scovillei, causal agents of pepper anthracnose, in potato dextrose agar (PDA) medium decreased the aggressiveness and virulence of the fungus in host‐pepper fruits. However, reintroduction of the pathogen to the host and isolation of the pathogen as the new inoculum, prior to inoculation assays, increased the virulence of the fungi. These findings suggest that re‐inoculation and re‐isolation of Colletotichum truncatum and C. scovillei that have been stored for at least 1 year in PDA medium are necessary when using fungal cultures in pathogenicity and plant resistance assays to achieve desirable, comparable and reliable results.     

Growth promotion effect of Pichia guilliermondii in chilli and biocontrol potential of Hanseniaspora uvarum against Colletotrichum capsici causing fruit rot

In the present investigation, we have attempted to identify the potential two epiphytic yeast strains for growth promotion and management of chilli fruit rot. Seed treatment with Pichia guilliermondii showed increased seedling vigour index (55%), fresh weight (96%) and dry weight (45%) over untreated control. Furthermore, P. guilliermondii showed higher root colonisation ability, indole-3-acetic acid (IAA) production and phosphate solubilisation ability. On the other hand, seedling dip with Hanseniaspora uvarum induced higher levels of defence-related compounds in chilli seedlings challenge-inoculated with Colletotrichum capsici under glasshouse conditions. Among the different media tested, higher biomass of P. guilliermondii and H. uvarum was obtained in pine juice broth and sugarcane juice broth, respectively. Glycerol buffer formulation showed viability (>70%) of P. guilliermondii up to 4 months and H. uvarum up to 9 months when stored at ambient conditions. Seedling dip and foliar sprays with H. uvarum showed 37– 40% reduction in chilli fruit rot incidence under field conditions. It also showed higher (cumulative) accumulation of defence-related compounds in chilli leaves and ripe fruits under field conditions. The results of current investigation indicated a clear difference among the two epiphytic yeast strains. P. guilliermondii was identified as growth promoter of chilli and H. uvarum as antagonist of chilli fruit rot pathogen, C. capsici.     

Development of CMV-and TMV-resistant chili pepper: field perfermance and biosafety assessment

Both cucumber mosaic virus (CMV) and tobacco mosaic virus (TMV) coat protein (CP) genes have been transferred to chilli pepper (Capsicum annuum var. Longunt) cultivar 8212 by a modified procedure of Agrobacterium tumefaciens-mediated transformation using hypocotyl as the explant. PCR analysis revealed the presence of both CMV and TMV CP genes in at least 11 primary transformants out of 49 kanamycin-resistant chili pepper plants. Ten T1 lines from five independent transformation events were identified as putative homozygous transgenic lines based on the rooting assay of their T2 seedlings on the kanamycin-containing media. Integration and expression of CMV CP and TMV CP transgenes in one of the homozygous line, 16-13, were confirmed bySouthern blot, RT-PCR and western blot analyses. Line 16-13 was highly resistant to infection of homologous CMV and TMV strains in greenhouse conditions when successively challenged with CMV and TMV or challenged with TMV alone.Futhermore, field trials on T2, T3 and T4 progenies of Line 16-13 were performed on scales of 123, 300 and 10,000 plants, respectively, in consecutive years 1996, 1997 and 1998 with the permission of the Chinese government authority. The transgenic plants displayed delayed symptom development and significantly milder disease severity in field conditions when compared to untransformed chili pepper plants, resulting in 47 and 110% increase in pepper fruit yield in surveys conducted in 1997 and 1998 trials, respectively. Finally, quality analysis and biosafety assesment were performed on transgenic chili pepper fruit concurrently with the control fruit, and demonstrated that the transgenic chili pepper fruit is substantially equivalent to the non-transgenic pepper in terms of the quality and biosafety when consumed as a food additive.     

Thidiazuron induced somatic embryogenesis and plant regeneration in Capsicum annuum

An efficient protocol of direct somatic embryogenesis (without involving intermediate callus) has been developed from stem segments and shoot tips of Capsicum annuum L. Explants were cultured on Murashige and Skoog (MS) medium supplemented with thidiazuron (TDZ). Among the various concentration of TDZ tested, 0.5 μM was proved to be best for induction of somatic embryos. Induction, maturation and germination were achieved on the same medium. The shoots developed from somatic embryos were transferred for rooting to MS medium supplemented with indole-3-butyric acid (IBA). All the regenerated plants with 85 % survival rate were normal with respect to morphology and growth characteristics.     

Screening of chilli germplasm for resistance to Alternaria leaf spot disease

Chilli is one of the spices used to enhance the flavour and taste of cooked food. Fungal diseases are the main biological constraints in chilli production, and Alternaria leaf spot disease caused by Alternaria alternata is one of the most devastating diseases of chilli. One of the effective and environmentally friendly ways to control this disease is introgress resistance from wild relative/varieties to the cultivated one. The first step towards introgression of resistance genes is to screen the chill germplasm for leaf spot resistance. In the current study, we screened the chilly germplasm and identified the sources of leaf spot resistance, which can be harnessed in resistance breeding programmes.     

Predators do not spill over from forest fragments to maize fields in a landscape mosaic in central Argentina

South America is undergoing a rapid and large‐scale conversion of natural habitats to cultivated land. Ecosystem services still remain important but their level and sustainability are not known. We quantified predation intensity in an Argentinian agricultural landscape containing remnants of the original chaco serrano forest using artificial sentinel prey. We sought to identify the main predators and the effect of landscape configuration and maize phenology on predation pressure by invertebrate and vertebrate predators in this landscape. The most common predators were chewing insects (50.4% predation events), birds (22.7%), and ants (17.5%). Overall predation rates in forest fragments (41.6% per day) were significantly higher than in the surrounding maize fields (21.5% per day). Invertebrate predation was higher inside and at the edge of forest fragments than within fields, and did not change with increasing distance from a fragment edge, indicating a lack of spillover from the native habitat remnants to the cultivated matrix at the local scale. Distance from a continuous forest had a positive impact on predation by invertebrates and a negative impact on vertebrate predation.     

Fruit characteristics, seed production and pollen tube growth in the wild chilli pepper Capsicum flexuosum

Fruits formed after different pollination regimes (flowers hand pollinated, unpollinated, and open pollinated) and the seeds obtained were characterized in the wild chilli pepper Capsicum flexuosum Sendtn. Pollen tube development in vivo and ovary growth were also analyzed. Seedless fruits and empty seeds were abundant among the fruits from hand pollinated and open pollinated flowers, while no more than one seed with embryo was found in a low percentage of fruits from such pollination treatments. Parthenocarpic fruits were formed from unpollinated flowers. Pollen tube growth was arrested in the upper third of the style for almost all pollen tubes except for a single one that may continue elongating occasionally. The ovary size increased continuously after pollination, even without fertilization. The sum of the evidence registered may help to explain the low number of seeds with embryo harvested, the abundance of seedless fruits formed from pollinated flowers (possibly parthenocarpic), and the high rate of parthenocarpic fruits formed from unpollinated flowers.     

Nucleotide sequence of the 3′-terminal region of chilli vein-banding mottle potyvirus isolated from pepper in Thailand

The sequence of the 2227 nucleotides of the 3-terminus of chilli vein-banding mottle virus-Chiangmai isolate (CVbMV-CM1) genome was determined. This sequence encodes a putative 287 amino acid coat protein. Downstream of coat protein coding region is a 288 nucleotide untranslated sequence terminated by a polyadenylate tract. The amino acid sequence of coat protein contained aspartic acid–alanine–glycine tripeptide, which may correlate with the high frequency of aphid transmissibility. The cleavage site between large nuclear inclusion protein (NIB) and coat protein was glutamine/serine. Comparison of 3 non-coding region nucleotide and coat protein amino acid sequences of CVbMV-CM1 with those of other 18 potyviruses showed 44–56% and 54–65% homology respectively, suggesting CVbMV should be regarded as new potyvirus.