Genetic variability in mulberry against Myrothecium leaf spot (Myrothecium roridum) and identification of resistance genotypes

Disease response of 30 mulberry genotypes to Myrothecium leaf spot (Myrothecium roridum) was studied under inoculated condition. It was observed that 10 genotypes were resistant, 16 genotypes moderately resistant and 4 genotypes moderately susceptible to the disease. Area under disease progress curve (AUDPC) and apparent infection rate was found significantly lower in the resistant genotypes. Correlation study revealed that percent disease index (PDI) has significant positive correlation with AUDPC and apparent infection rate. Genetic analysis of disease-resistant traits (PDI and mean AUDPC) revealed that phenotypic coefficient of variation (PCV) was higher than genotypic coefficient of variation (GCV) for both PDI and AUDPC, and GCV/PCV ratio was also found high which indicated that disease-resistant traits to Myrothecium leaf spot were not much influenced by environment. High heritability coupled with high genetic advance indicated that the disease-resistant traits are due to additive gene effect thereby indicating the amenability of disease-resistant in the selection process. Hence disease-resistant mulberry genotypes viz. C-763, S-34, Jodhpur, Cyprus, Australian and Hungarian may be used as source of resistance to Myrothecium leaf spot for future breeding programme. Besides, high yielding genotypes viz. Tr-10, C-763 and S-34 may be recommended for commercial exploitation     

Screening of mulberry germplasm lines against Powdery mildew,Myrothecium leaf spot and Pseudocercospora leaf spot disease complex

Abstract

Fifty-six indigenous and 29 exotic mulberry varieties were screened against powdery mildew, Myrothecium leaf spot, Pseudocercospora leaf spot for a period of three years under field conditions. The percent disease index (PDI) was recorded during the peak season of the diseases. Out of 85 germplasm lines studied four-germplasm lines viz. Thailand lobed (0.43), M. multicaulis (2.63), Italian (2.83) and M. australis (4.56) were found highly resistant; nine lines were resistant; 43 lines were moderately resistant and 29 were susceptible to the disease complex. Powdery mildew showed significant positive correlation with Pseudocercospora leaf spot. Highly resistant varieties may be utilized for future disease resistance breeding programme to evolve multiple disease resistant mulberry varieties.     

Response of mulberry accessions to major foliar diseases in temperate conditions of Jammu and Kashmir,India

Thirty-three indigenous and 24 exotic mulberry accessions belonging to five Morus spp. originated from seven countries distributed in temperate and tropical climates were observed for their response to two major foliar diseases during 2010 and 2011 under temperate conditions of Jammu and Kashmir, India. Leaf spot and powdery mildew severity (Percent Disease Index (PDI)) ranged from 0.00 to 74.90% and 0.00 to 59.85%, respectively. Indigenous and exotic accessions responded similarly to leaf spot, but varied too much to powdery mildew. Irrespective of origin, response in ascending order of PDI for leaf spot is M. multicaulis, M. indica, M. alba, M. kayayama and M. bombycis and for powdery mildew is M. multicaulis, M. kayayama, M. bombycis M. alba and M. indica. Among indigenous accessions, Brentul Kashmir offered highest resistance to both the diseases. Nadigam offered maximum resistance only to leaf spot followed by Himachal local; while Chinarpati followed by Mysore local only for powdery mildew. Among exotic accessions, Ichinose offered maximum resistance to both the diseases followed by Kokusou-21 and Tagowase.     

Arachidonic acid-induced hypersensitive cell death as an assay of downy mildew resistance in pearl millet

Arachidonic acid (AA) induces hypersensitive response (HR) on coleoptile/root regions of two-day-old pearl millet seedlings. The response is comparable to the HR induced by the downy mildew pathogen, Sclerospora graminicola. A time gap in the appearance of cell necrosis among genotypes of pearl millet was related to the degree of resistance to downy mildew. Based on the time required for the development of necrotic spots induced by AA, the pearl millet genotypes were categorised as highly resistant/resistant (HR in 3–6 h), susceptible (HR in 7–12 h) and highly susceptible (HR in 13 h and above). The percentage disease incidence in each genotype was compared with the time required for the development of AA-induced HR. The appearance of hypersensitive cell necrosis was rapid in genotypes having high resistance to downy mildew and was slow in genotypes with high susceptibility. This simple method of screening various pearl millet genotypes in the absence of the pathogen aids in identifying the downy mildew resistant/susceptible host cultivars without the risk of introducing the virulent race of the pathogen.     

Tropic failure of Phyllactinia corylea contributes to the mildew resistance of mulberry genotypes

Different mulberry genotypes show great variation in their resistance to the powdery mildew Phyllactinia corylea. Conidial germination and hyphal growth of P. corylea on the leaf surface of two susceptible mulberry genotypes, viz., Kanva 2 (K2) and Victory 1 (V1) varieties of Morus indica, and on two resistant species, viz., M. laevigata and M. serrata were studied by scanning electron microscopy. Conidial germination and growth of germ tubes were normal on all the leaves. The hyphae of P. coryleaidentify stomata on host leaves by their topographical features to produce the stomatopodia precisely over them. The holes and/or the grooves of stomata appear to provide the signals for the initiation of stomatopodia and similar structures are erratically developed over many local depressions or grooves on leaf surface. The abaxial surface of K2 leaf is smooth without prominent undulations of epidermal cell surface, and the stomata are flush with the leaf surface. Although successful penetration is also achieved on V1 leaf, its slightly undulated surface occasionally provides inaccurate tropic signals to the hyphae, inducing the development of stomatopodia away from the stomata. The leaf surfaces of M. laevigata and M. serrata are very rough with highly sculptured cuticle and abundant epidermal outgrowths. Stomata mostly remain sunken or hidden amidst the cuticular ornamentations and the hyphae fail to recognise the precise signals from them. As the surface architecture of the leaves provides many immense sources of tropic signals, stomatopodia are often produced over local depressions or grooves. In these cases the fungus fails to penetrate the leaf, does not develop beyond 24 h and penetration is rarely achieved on the leaves of the resistant plants. The study indicates that the stimulatory effect of the leaf surface topography of resistant varieties misleads the pathogen from successful penetration, thus contributing to the plant's resistance.This revised version was published online in October 2005 with corrections to the Cover Date.     

Genetic relationships of Japanese and Indian mulberry (Morus spp.) genotypes revealed by DNA fingerprinting

Mulberry (Morus spp.), a deciduous tree, originated at the foothills of the Himalayas and is used in sericulture for its leaf to feed the silkworm Bombyx mori L. Species differentiation among the genotypes of the genus Morus has never been out of debate as inter-specific hybridization events are often fertile. In the present study attempts were made to elucidate the genetic relationships among 18 mulberry genotypes collected from India and Japan using 15 Inter Simple Sequence Repeat (ISSR) and 15 Random Amplified Polymorphic DNA (RAPD) primers. The ISSR primers generated 81.13% polymorphism while the RAPDs generated 71.78% polymorphism. The polymorphic index of the primers identified UBC-812, UBC-826, UBC827, UBC-881, OPA-01, OPA-02, OPA-04 and OPH-17 as informative primers in mulberry. The genetic similarity coefficients and the dendrograms showed considerable genetic similarity among the genotypes. However, using the DNA markers, these genotypes were discriminated into two major groups in accordance with their geographic origin and species status. Distribution of the genotypes on a two-dimensional figure on the basis of the ALSCAL algorithm using Euclidean distance further confirmed the genetic divergence between these two groups. From the study it can be concluded that though morphologically Japanese and Indian mulberry genotypes show little divergence, genetic analysis using DNA markers could unravel significant genetic variation between these two groups. Similarly, while the species status of Japanese mulberry genotypes agrees with the genetic analysis, the same does not apply to Indian genotypes, in agreement with many earlier reports. The information generated in this study is of much use for taxonomical grouping and also for utilization in breeding and conservation programs.     

A novel strain of Pseudozyma aphidis from mulberry parasitises the conidia of mulberry powdery mildew fungus Phyllactinia sp. and its biocontrol effect in the fields

Plant surface is colonised with a vast community of non-pathogenic epiphytic microorganisms which play an important role in host defence. In the present study, we reported a fungus from mulberry leaf surface that showed an antagonistic effect against mulberry powdery mildew fungal pathogen Phyllactinia sp. This novel isolate is a yeast-like fungus that was identified as Pseudozyma aphidis CNm2012 based on morphologic and phylogenetic analysis. According to our research, P. aphidis CNm2012 directly acted on the powdery mildew conidia via parasitism which caused conidial atrophy, collapse and eventually, cleavage and death. During the parasitic process, we found the isolate could gather around the conidia of Phyllactinia sp. and form hyphae that grew on the conidial surface and utilise the conidia as a nutrient source. Field studies revealed that P. aphidis CNm2012 could suppress the disease incidence of mulberry powdery mildew caused by Phyllactinia sp., and reduce the disease severity. To our knowledge, it is the first report of P. aphidis directly against powdery mildew fungus Phyllactinia spp. by parasitism. Our results indicated that P. aphidis CNm2012 could be served as an environmentally friendly alternative of chemical pesticides to manage mulberry powdery mildew disease.     

Breeding Potential of Some Exotic Tomato Lines: A Combined Study of Morphological Variability,Genetic Divergence,and Association of Traits

Tomato (Solanum lycopersicum L.) is called ‘the poor man’s orange’ due to its low price and improved nutritional values. An experiment was conducted to study the breeding potential of some exotic tomato lines by assessing various qualitative and quantitative traits conferring yield and quality attributes. Among the qualitative traits, greater variability was observed for growth type, stem hairiness, and fruit shape and size. A determinate growth habit was observed in the genotype AVTO9802 while the genotype AVTO0102 produced yellow color fruits. A significant (p ≤ 0.01) variation was also observed for the studied quantitative traits. Based on yield and traits attributed to yield, the genotypes AVTO0314, GPB0107, GPB0120 and AVTO9802 were selected as promising genotypes. The differences between the genotypic and phenotypic coefficients of variation (GCV and PCV) of the studied quantitative traits were very low. This suggests that the apparent variation was mainly due to the genotypes. The higher GCV and PCV values were observed for the number of primary branches plant⁻¹ (NPB), number of fruits cluster⁻¹ (NFC), individual fruit weight (IFW) and total soluble solids (TSS). High heritability was recorded for all quantitative traits in a broad sense. However, the individual fruit diameter showed the highest heritability (99.56). The highest (102.75) genetic advance (GA) was observed for the number of fruits plant⁻¹ (NFP). High heritability coupled with high GA as percentage of mean were recorded for the traits NFP, NFC, fruit yield plant⁻¹ (FYP) and IFW. FYP showed a significant positive correlation with NFC (0.714***) and a negative correlation with days to the first harvest (−0.539***) and plant height (−0.492**). Principal component analysis revealed that the first four components explained 78.5% of the total variation among the genotypes. Thus, the promising genotypes (AVTO0314, GPB0107, GPB0120, AVTO9802 and AVTO0102) isolated from this study can be used for developing high-yielding and high-quality tomato varieties.     

Evaluation of Gossypium species for resistance to cotton leaf curl Burewala virus

Cotton leaf curl disease (CLCuD), caused by cotton leaf curl Burewala virus (CLCuBV), has emerged as a major threat to cotton production in Pakistan. Resistance to CLCuBV was evaluated in cultivated and wild cotton genotypes representing six Gossypium species by visual symptom scoring and virus assessment using PCR tests. Considerable variation in responses was observed when using whitefly and graft transmission to inoculate Gossypium genotypes with CLCuBV in field and greenhouse experiments. Under field evaluation, all cultivated genotypes of Gossypium hirsutum and three genotypes of G. barbadense were susceptible. Eleven genotypes that represented six wild and cultivated Gossypium species were considered to be highly resistant as they were free from infection. Similar results were obtained when these genotypes were tested using whitefly transmission. To verify these findings, 132 cultivated and wild genotypes were tested by graft inoculation. All G. hirsutum genotypes (116 cultivated, 1 wild, 1 transgenic Coker-312 and 1 non-transgenic Coker-312), three G. barbadense genotypes and one G. thurberi genotype were highly susceptible and exhibited symptoms 9–12 days after grafting. Four genotypes of G. arboreum and one genotype of G. anomalum did not express symptoms but had a detectable level of virus. One genotype of G. herbaceum and three wild genotypes of G. hirsutum showed mild symptoms (severity indexes of 1–2) and exhibited delayed disease development. These genotypes were classified as moderately resistant to resistant. Resistant genotypes that were identified in this study will be useful sources for exploitation of breeding programmes aimed at developing CLCuBV-resistant varieties and increasing genetic diversity.     

A comparison of genetic variation among wild and cultivated Morus Species (Moraceae: Morus) as revealed by ISSR and SSR markers

In this study, inter-simple sequence repeats (ISSR) ans simple sequence repeat (SSR) markers were used to investigate genetic diversity of 27 mulberry accessions including 19 cultivated accessions (six M. multicaulis, three M. alba, two M. atropurpurea, two M. bombycis, one M. australis, two M. rotundiloba, one M. alba var. pendula, one M. alba var. macrophylla, and one M. alba var. venose) and 8 wild accessions (two M. cathayana, two M. laevigata, two M. wittiorum, one M. nigra and one M. mongolica). ISSRs and SSRs were compared in terms of their informativeness and efficiency in a study of genetic diversity and relationships among 27 mulberry genotypes. SSRs presented a higher level of polymorphism and greater information content. All index values of genetic diversity both markers analyzed using Popgene 32 software indicated that within wild species had higher genetic diversity than within cultivated species. Cultivation may caused the lose of genetic diversity of mulberry compared with wild species revealed by ISSR and SSR markers. The mean genetic similarity coefficients among all mulberry genotypes ascribed by ISSR and SSR matrices were 0.7677 and 0.6131, respectively. For all markers a high similarity in dendrogram topologies was obtained although some differences were observed. Cluster analysis of ISSR and SSR using UPGMA method revealed that the wild species are genetically distant from the domesticated species studied here. The correlation coefficients of similarity were statistically significant for both marker systems used. Principal coordinates analysis (PCA) for ISSR and SSR data also supports their UPGMA clustering. These results have an important implication for mulberry germplasm characterization, improvement, molecular systematics and conservation.     

Tropic failure of Phyllactinia corylea contributes to the mildew resistance of mulberry genotypes

Different mulberry genotypes show great variation in their resistance to the powdery mildew Phyllactinia corylea. Conidial germination and hyphal growth of P. corylea on the leaf surface of two susceptible mulberry genotypes, viz., Kanva 2 (K2) and Victory 1 (V1) varieties of Morus indica, and on two resistant species, viz,, M. laevigata and M. serrata were studied by scanning electron microscopy. Conidial germination and growth of germ tubes were normal on all the leaves. The hyphae of P. corylea identify stomata on host leaves by their topographical features to produce the stomatopodia precisely over them. The holes and/or the grooves of stomata appear to provide the signals for the initiation of stomatopodia and similar structures are erratically developed over many local depressions or grooves on leaf surface. The abaxial surface of K2 leaf is smooth without prominent undulations of epidermal cell surface, and the stomata are flush with the leaf surface. Although successful penetration is also achieved on V1 leaf, its slightly undulated surface occasionally provides inaccurate tropic signals to the hyphae, inducing the development of stomatopodia away from the stomata. The leaf surfaces of M. laevigata and M. serrata are very rough with highly sculptured cuticle and abundant epidermal outgrowths. Stomata mostly remain sunken or hidden amidst the cuticular ornamentations and the hyphae fail to recognise the precise signals from them. As the surface architecture of the leaves provides many immense sources of tropic signals, stomatopodia are often produced over local depressions or grooves. In these cases the fungus fails to penetrate the leaf, does not develop beyond 24 h and penetration is rarely achieved on the leaves of the resistant plants. The study indicates that the stimulatory effect of the leaf surface topography of resistant varieties misleads the pathogen from successful penetration, thus contributing to the plant's resistance.     

Assessment of sorghum genetic resources of Ethiopia for anthracnose (Colletotrichum sublineolum Henn.) resistance and agronomic traits

Sorghum anthracnose caused by Colletotrichum sublineolum Henn. is one of the key diseases limiting sorghum production and productivity. Development of anthracnose‐resistant sorghum genotypes possessing yield‐promoting agronomic traits is an important breeding goal in sorghum improvement programs. The objective of this study was to determine the responses of diverse sorghum genetic resources for anthracnose resistance and agronomic traits to identify desirable lines for breeding. A total of 366 sorghum collections and three standard checks were field evaluated during the 2016 and 2017 cropping seasons. Lines were artificially inoculated with a virulent pure isolate of the pathogen. Anthracnose disease severity was assessed to calculate the area under disease progress curve (AUDPC). Agronomic traits such as panicle length (PL), panicle width (PW), head weight (HW) and thousand grain weight (TGW) were measured. Lines showed highly significant differences (p < .001) for anthracnose severity, AUDPC and agronomic traits. Among the collections 32 lines developed levels of disease severity between 15% and 30% in both seasons. The following sorghum landraces were selected: 71708, 210903, 74222, 73955, 74685, 74670, 74656, 74183, 234112, 69412, 226057, 214852, 71420, 71484, 200126, 71557, 75120, 71547, 220014, 228179, 16212, 16173, 16133, 69088, 238388, 16168 and 71570. These landraces had a relatively low anthracnose severity possessing farmer‐preferred agronomic traits. The selected genotypes are useful genetic resources to develop anthracnose‐resistant sorghum cultivars.     

Somaclonal variation in embryogenic cultures of silver fir (Abies alba Mill.)

Abstract

The somaclonal variation analysis was conducted on callus of 57 lines obtained by the method of somatic embryogenesis from six zygotic embryos (with different genotypes) of silver fir (Abies alba Mill.) located in two mountain regions in the south of Poland. The somaclonal variation at the DNA level was estimated using RAPD markers and the data produced were used to estimate the level of similarity using Jaccard’s coefficient. For RAPD analysis, 24 ten‐nucleotide primers from the groups OPA, OPB and OPG were used. Two genotypes deriving from Krynica and My?lenice showed high genetic similarity (Jaccard’s coefficient 0.74 and 0.83), which provides a substantial chance for producing firs with the parental genotype. The remaining four genotypes showed somaclonal variation (average Jaccard’s coefficient approx. 0.5). The significance in variation of the research sites was ascertained by the ANOVA statistical test, which showed the impact of genotype, type of medium and phytohormones included in it on the variation among the fir lines bred in vitro. The somaclonal variation data in silver fir could be useful for its propagation through in vitro culture, and in generating detailed genetic maps of this species.     

Occurrence of Hop Stunt Viroid in Mulberry (Morus alba) in Lebanon and Italy

The presence of Hop stunt viroid (HSVd) was detected using RT‐PCR and Northern blot hybridization in five of 60 samples from symptomless mulberry trees (Morus alba) collected in Italian and Lebanese orchards in July 2010. Infection levels were c. 10% in Lebanese and 8% in Italian samples. Nucleotide alignments showed that sequences of the mulberry HSVd isolates shared 95–96% identity with those of the same viroid occurring elsewhere. In a phylogenetic tree, mulberry HSVd isolates clustered together with those of HSVd‐citrus, regardless of their geographical origin. This is the first report of infection in mulberry trees by HSVd.     

Sclerospora graminicola- and arachidonic acid-induced autofluorescence in downy mildew resistant and susceptible genotypes of pearl millet

Autofluorescence of downy mildew resistant and susceptible cells of pearl millet seedlings undergoing hypersensitive reaction (HR) upon Sclerospora graminicola-inoculation and arachidonic acid (AA)-treatment was studied. Two-day-old seedlings of a highly resistant (IP 18296) and a highly susceptible (23D₂B) genotype of pearl millet were either inoculated with zoospore suspension of S. graminicola or treated with AA for 24 h. The coleoptiles with hypersensitive necrotic spots were processed by the standard procedure, and the tissues were subjected to fluorescence microscopy. A differential accumulation of autofluor-escent compounds in resistant and susceptible pearl millet genotypes was observed with most accumulation occurring in resistant cells treated with AA. The variation in the degree of fluorescence and the spatial accumulation of autofluorescent compounds among the two inoculated/treated genotypes is discussed.     

Molecular characterization of urdbean (Vigna mungo) germplasm related to resistance against urdbean leaf crinkle virus

Urdbean (Vigna mungo) is an important pulse crop grown worldwide. Urdbean leaf crinkle virus (ULCV) is a pathogen of urdbean found in Pakistan that causes huge losses in yield. Forty urdbean varieties/lines were screened against the virus under field conditions during spring season 2009. None of the lines appeared to be highly resistant or resistant. On the basis of a 0-5 disease rating scale and disease severity index, genotypes varied significantly in their reaction to ULCV. Four lines (M-6206, IAM-382-15, IAM-133, and Mash-1) were moderately resistant, eight were rated as moderately susceptible, and 21 as susceptible; the remaining seven lines were highly susceptible. RAPD analyses revealed an extensive amount of variation, which could be used for cultivar identification. Genetic differentiation among urdbean genotypes was similar to the field screening data. The varieties 6065-3 and 6206 were highly susceptible and moderately resistant, respectively, to ULCV under field conditions, confirmed by the RAPD analysis. These varieties were the most diverse varieties in the similarity matrix (67.2%), while the varieties IAM-382-9 and 07M003 were the most similar (98.4%). This information will help in the recognition of available resistant germplasms that can resist this disease and will be utilized for urdbean improvement in Pakistan.     

Patterns of resistance of Israeli wild emmer wheat to pathogens I. Predictive method by ecology and allozyme genotypes for powdery mildew and leaf rust

The association of ecological factors and allozyme markers with genotypes of tetraploid wild emmer wheat,Triticum dicoccoides, varying in resistance to four cultures of the pathogenErysiphe graminis tritici, and to one culture ofPuccinia recondita tritici, which incite the diseases powdery mildew and leaf rust respectively, were explored theoretically and practically. The study involved 233 accessions comprising 10 populations representing the ecological range ofT. diococcoides in Israel. Our results indicate that genetic polymorphism for resistance to both pathogens is structured geographically, and is predictable by climatic as well as allozymmc markers. Three variable combinations of water factors and temperature differentials significantly explain 0.27 and 0.14 of the spatial variance for resistance to powdery mildew and leaf rust, respectively, suggesting the involvement of natural selection. Several allozyme genotypes, singly, or in combination, are significantly associated with disease resistance. We conclude thatT. dicoccoides populations in Israel, which grow in the center of diversity of the species, contain large amounts of unexploited disease resistant genotypes. The populations could be effectively screened and utilized for producing resistant cultivars by means of ecological factors and allozyme markers as predictive guidelines.