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Open Access
GENE ACTION FOR YIELD AND RELATED TRAITS IN SOYBEAN [Glycine max (L.) Merrill] AND DEVELOPMENT OF INTERSPECIFIC HYBRIDS INVOLVING WILD SPECIES
(CSKHPKV Palampur, 2014-12) INDU BALA; Jai Dev
The present investigation entitled “Gene action for yield and related traits in soybean [Glycine max (L.) Merrill] and development of interspecific hybrids involving wild species” was carried out at the Experimental Farm of the Department of Crop Improvement, CSK HPKV, Palampur to gather information on genetic architecture for seed yield and component traits in soybean and to introgress desirable genes from wild species to cultivated ones. The experimental material comprised of 54 triple test cross progenies derived by mating 18 lines with three testers, namely, Hara Soya (L1), Him Soya (L2) and their F1 (L3). This genetic material was evaluated in a randomized complete block design with three replications during kharif 2013. Epistasis was found to be an integral part of genetic variation for majority of the traits. Epistatic interaction for many traits was ‘j+l’ type except days to 50% flowering, days to 75% maturity, reproductive phase, petiole length, seed per pod, harvest index and 100 seed weight, whereas plant height, internode length, pods per plant and biological yield per plant carried ‘i’ type epistasis alongwith ‘j+l’ type. Additive component (D) was more pronounced than dominance component (H) for most of the traits. Both additive and dominance components were of almost equal magnitude for pod length indicating the importance of both additive and dominance type of gene action, whereas partial degree of dominance was noticed for majority of traits. The kind of genetic variance revealed from triple test cross can be exploited by intermating selected individuals in early segregating generations with delayed selection in later generations, diallel selective mating/ biparental mating or recurrent selection followed by pedigree method to exploit both additive and non-additive components alongwith epistasis. Lines Bragg, Shivalik and P9-2-2 were found to be good general combiners for most of the traits. The cross combinations, Bragg x Hara Soya, PK- 472 x Him Soya, DS-1213 x Hara Soya, H-330 x Hara Soya and H-330 x Him soya showed high per se performance, SCA effects, heterobeltiosis and economic heterosis for seed yield per plant and were rated as potential crosses for further improvement. Cross H-330 x Him Soya showed resistance against brown spot and bacterial pustule diseases. The interspecific hybrids developed involving Glycine max x G. soja, were true to type based on confirmation at the morphological, molecular and cytological level. Sufficient variability was found for all the traits studied in wide hybrids. Seed yield per plant showed significantly positive correlation with reproductive phase, pods per plant, pod length, petiole length, harvest index and 100-seed weight.
Open Access
Variability, epidemiology and management of Fusarium oxysporum f.sp. lini.
(CSK HPKV, Palampur, 2019-06) Narinder Pal; Ashok Kumar
The investigation entitled “Variability, epidemiology and management of Fusarium oxysporum f.sp. lini” was conducted at Department of Plant Pathology, CSKHPKV Palampur and Shivalik Agricultural Research and Extension Centre, Kangra during 2015-2018. 35 isolates of Fusarium oxysporum f.sp. lini were obtained from the diseased samples collected from Kangra and Mandi districts of Himachal Pradesh and other parts of India. Three isolates were categorized as weakly pathogenic, 7 as moderately pathogenic and 25 as highly pathogenic during pathogenicity test. All the isolates showed variability in morphological and cultural characteristics. To identify the wilt resistant sources, 176 linseed genotypes were evaluated under field conditions. Among these 5 genotypes viz., KL-215, KL-261, EC541213, H-18 and TL-50 showed highly resistant,7 genotypes showed resistant and 21 genotypes showed moderately resistant disease reactions. A set of 17 genotypes i.e. TL-9, R-552, H-34, Janaki, Himalini, Polf-24, TL-22, EC-541213, KL-187, POLF-16, EC-541212, H-18, EC-541211, POLF-22, Kangra local, Baner and Chambal was standardized as differential hosts. Pathogenic variability among 35 isolates was determined on differential set and 5 pathotypes were recorded and designated as Pathotype-1 to Pathotype-5. 32 promising genotypes of linseed were evaluated against the 5 pathotypes and 13 genotypes viz., EC-541199, H-18, H-36, Him Alsi-1, Jeevan, Janaki, KL-215, KL-227, KL-265, Nagarkot, Natazo, POLF-19 & Surbhi showed resistant disease reaction against all the 5 pathotypes. Maximum in vitro growth of the Fusarium oxysporum f.sp. lini on PDA was observed at 24ºC and pH 5.5 with highest growth rate and sporulation. Lowest wilt incidence was recorded in clay loam soil compared to sandy, sandy clay loam and loam soils. Disease incidence during the crop season 2015-16 and 2016-17 showed positive correlation with minimum temperature and soil temperature at 10 cm and negative correlation with relative humidity at evening time. Delay in date of sowing from 16 October to 26 November resulted decrease in wilt incidence in varieties Chambal and Janaki. Trichoderma viride (Strain Tr-3) & Carbendazim 50 WP were found most effective for mycelial growth inhibition of the Fusarium oxysporum f.sp. lini during in vitro studies and Tr-3 (Trichoderma viride) showed highest compatibility with fungicides among three tested bioagents. In pot studies, the highest seedling emergence and lowest linseed wilt incidence was observed with Tr-4 (Trichoderma harzianum) (seed treatment + soil application), Thiram 75WP seed treatment and Thiram 75 WP + Tr-4 (Trichoderma harzianum) (seed treatment + soil application). In field studies, Carbendazim (50%) + Tr-4 (T. harzianum) (seed treatment + soil application) showed highest disease reduction and yield increase over control.
Open Access
VARIABILITY, EPIDEMIOLOGY AND MANAGEMENT OF Fusarium oxysporum f.sp. lini.
(CSKHPKV, Palampur, 2019-06) Pal, Narinder; Ashok Kumar
The investigation entitled “Variability, epidemiology and management of Fusarium oxysporum f.sp. lini” was conducted at Department of Plant Pathology, CSKHPKV Palampur and Shivalik Agricultural Research and Extension Centre, Kangra during 2015-2018. 35 isolates of Fusarium oxysporum f.sp. lini were obtained from the diseased samples collected from Kangra and Mandi districts of Himachal Pradesh and other parts of India. Three isolates were categorized as weakly pathogenic, 7 as moderately pathogenic and 25 as highly pathogenic during pathogenicity test. All the isolates showed variability in morphological and cultural characteristics. To identify the wilt resistant sources, 176 linseed genotypes were evaluated under field conditions. Among these 5 genotypes viz., KL-215, KL-261, EC541213, H-18 and TL-50 showed highly resistant,7 genotypes showed resistant and 21 genotypes showed moderately resistant disease reactions. A set of 17 genotypes i.e. TL-9, R-552, H-34, Janaki, Himalini, Polf-24, TL-22, EC-541213, KL-187, POLF-16, EC-541212, H-18, EC-541211, POLF-22, Kangra local, Baner and Chambal was standardized as differential hosts. Pathogenic variability among 35 isolates was determined on differential set and 5 pathotypes were recorded and designated as Pathotype-1 to Pathotype-5. 32 promising genotypes of linseed were evaluated against the 5 pathotypes and 13 genotypes viz., EC-541199, H-18, H-36, Him Alsi-1, Jeevan, Janaki, KL-215, KL-227, KL-265, Nagarkot, Natazo, POLF-19 & Surbhi showed resistant disease reaction against all the 5 pathotypes. Maximum in vitro growth of the Fusarium oxysporum f.sp. lini on PDA was observed at 24ºC and pH 5.5 with highest growth rate and sporulation. Lowest wilt incidence was recorded in clay loam soil compared to sandy, sandy clay loam and loam soils. Disease incidence during the crop season 2015-16 and 2016-17 showed positive correlation with minimum temperature and soil temperature at 10 cm and negative correlation with relative humidity at evening time. Delay in date of sowing from 16 October to 26 November resulted decrease in wilt incidence in varieties Chambal and Janaki. Trichoderma viride (Strain Tr-3) & Carbendazim 50 WP were found most effective for mycelial growth inhibition of the Fusarium oxysporum f.sp. lini during in vitro studies and Tr-3 (Trichoderma viride) showed highest compatibility with fungicides among three tested bioagents. In pot studies, the highest seedling emergence and lowest linseed wilt incidence was observed with Tr-4 (Trichoderma harzianum) (seed treatment + soil application), Thiram 75WP seed treatment and Thiram 75 WP + Tr-4 (Trichoderma harzianum) (seed treatment + soil application). In field studies, Carbendazim (50%) + Tr-4 (T. harzianum) (seed treatment + soil application) showed highest disease reduction and yield increase over control.
Open Access
STUDIES ON GENE ACTION FOR SEED YIELD AND ITS RELATED TRAITS IN URDBEAN [Vigna mungo (L.) Hepper]
(CSKHPKV, Palampur, 2018-09-18) Patial, Ranjana; Mittal, R.K.
The present investigation entitled “Studies on gene action for seed yield and its related traits in urdbean [Vigna mungo (L.) Hepper]” was carried out at the Experimental Farm of the Department of Crop Improvement, CSK HPKV, Palampur to gather information on genetic architecture for seed yield and component traits in urdbean. The experimental material comprised of 81 triple test cross progenies developed by mating 27 lines with three testers viz., HPBU-111 (L1), Him Mash-1(L2) and F1 of HPBU-111and Him Mash-1 (L3). These F1’s along with their parents were raised in randomized block design with three replications during Kharif 2017. Data were recorded and analysed for 12 agro-morphological traits. Sufficient variability was observed in the TTC progenies for all the traits studied. Epistasis was found to be an integral part of genetic variation for all of the traits under study. ‘i’ type epistatic interaction estimates were significant for days to 50% flowering, days to 75% maturity, branches per plant, pods per plant, seed yield per plant, biological yield per plant, harvest index and 100 seed weight. Whereas, ‘j+l’ type interaction was significant for all the traits. Both additive and dominant components were significant for all the traits; where additive and dominance components were of almost equal magnitude for pod length, seeds per pod and 100 seed weight indicating the importance of both additive and dominance type of gene action and additive component being more pronounced for rest of the traits indicating the relative importance of fixable type of gene action in their inheritance. Combining ability estimates showed significant genetic variance in lines for all traits whereas testers had significant genetic variance for nine traits. Lines IC-436910, IC-413306, IC-IC-398973 and IC-343885 were found to be good general combiners for most of the traits. Crosses IC-343885 x HPBU-111 (G x G), IC-436910 x HPBU-111 (G x G), IC-413306 x Him Mash-1 (G x P) and IC-343943 x HPBU-111(P x G) were potential crosses on the basis of SCA estimates, mid parent heterosis, better parent heterosis and standard heterosis for seed yield and most of the traits. With regards to the Comparison of ТГС and L x T, the former is advantageous in providing an unambiguous test for the presence of epistasis, while the latter provides additional information, particularly with regard to the gca and sca effects and variances, helping breeders in the choice of better parents. Hence, both these designs should be applied together in order to have concrete information about the genetic architecture of economic traits in any crop.
Open Access
CHARACTERIZATION OF PEPPER MILD MOTTLE VIRUS STRAINS AND EVALUATION OF RESISTANCE IN CAPSICUM
(CSKHPKV, Palampur, 2018-06) Nidhi Kumari; Sharma, P.N.
Capsicum (Capsicum annuum L. var.grossum Sendt) is an important spice and vegetable crop being cultivated worldwide. More than 20 viruses are known to infect Capsicum spp. across the world and Pepper mild mottle virus (PMMoV), a member of Virgaviridae family and Tobamovirus genus is emerging as a great threat to the capsicum cultivation both in protected and open conditions in Himachal Pradesh (H.P.). The present study on PMMoV was undertaken to identify the pathotype of PMMoV prevalent in H.P., production of polyclonal antiserum against PMMoV-CP expressed in E. coli, evaluation of resistance against PMMoV in capsicum and identification of attenuated/mild strain if any. Surveys conducted to assess the prevalence of mild mottle disease showed wide occurrence of the disease and out of 97 samples collected during surveys, 54 showed the presence of PMMoV in DAS-ELISA with maximum percentage of positive samples from district Kullu (88.89%) followed by Mandi (78.57%) district. The presence of PMMoV was confirmed through RT-PCR using coat protein (CP) specific primers where positive samples yielded amplification of ~743 bp. Isolates were selected for varaiblity assays on differential varieties of capsicum and CP gene sequence analysis. All the isolates produced symptoms like mosaic, color variations, leaf cupping, vein banding on susceptible cultivar California Wonder. Based on the pathogenic reaction on differential varieties and amino acid sequence of CP gene, all the test isolates were grouped and identified as pathotype P12 which can overcome L+, L1 and L2 resistance alleles. For production of polyclonal antiserum, the PMMoV-CP was over-expressed in E. coli using IPTG at 1mM final concentration with overnight incubation in shaking incubator at 16oC which resulted in induction of target recombinant protein with molecular weight ~26kDa. The antiserum generated through out sourcing, evaluated for its sensitivity and specificity through Western blot and DAC-ELISA. In Western blot assay, the test antiserum reacted strongly both with PMMoV-CP in purified protein and native CP in crude sap from PMMoV infected pepper plants, whereas no reaction was observed with healthy plant sap. In DAC-ELISA antiserum dilution up to 1:1000 was capable of differentiating the PMMoV infected sample from healthy samples. The antiserum did not react with other capsicum viruses viz., Tobacco mosaic virus (TMV), Cucumber mosaic virus (CMV), Tomato spotted wilt virus (TSWV), Pepper veinal mottle virus (PVMoV), Potato virus Y (PVY) and Tomato yellow leaf curl virus (TYLCV) antigen. Only two exotic capsicum accessions PI-159236 and PI-260429 were found resistant to PMMoV. None of the isolates showed the characteristics of attenuated/mild strain as all the isolates produced prominent symptoms on susceptible cv- California Wonder.
Open Access
VIRULENCE ANALYSIS OF Colletotrichum lindemuthianum AND ITS MANAGEMENT IN COMMON BEAN
(CSK,HPKV,PALAMPUR, 2017-07) Sharma, NAIYA; Sharma, P.N.
ABSTRACT This study was aimed to determine pathogenic and molecular variability in Colletotrichum lindemuthianum, the casual organism of bean anthracnose; to evaluate bean germplasm to find out the sources of resistance against prevalent races; validation of R genes using molecular markers; and to identify suitable fungicide, biocontrol agents and botanicals for the management of disease. Virulence spectrum of 65 isolates determined on a set of 12 bean differential cultivars revealed the existence of 27 races in North Western Himalayas. Race 503 contained maximum number of 12 isolates from diverse geographic regions. Fifteen races viz; 5, 6, 7, 16, 18, 51, 87, 99, 145, 179, 211, 259, 337, 503 and 1395 were identified for the first time as none of them resembled with previously known races in Himachal Pradesh hence increasing the total number to 44 in the state. The isolates from J&K (3) and Uttrakhand (13) categorized into different races constitute their first record from these two states. The virulence analysis suggested 4 interaction types in accordance with the genetic origin of differential cultivars that were infected by a particular race. Twenty-one races having virulence for genotypes of both the gene pools showed Type IV interaction. Nine of 12 differential cultivars were found to be infected by one isolate suggesting 9 virulence factors among 65 isolates of C. lindemuthianum. RAPD data at 43 per cent similarity coefficient distinguished various isolates into 8 clusters, whereas, ISSR markers categorized test isolates into 5 clusters using 43 per cent similarity coefficient as a cutoff point. There was no congruence between the pathogenic variability and the molecular diversity data, showing no utility of such markers in differentiation of the physiological races whose identity is based on pathogenic behavior of the given isolate involving many pathogenicity factors. Out of 313 accessions, 45 were found resistant to race 0 (20), 17 (21), 503 (19) and 1395 (13), respectively. R-gene prediction analysis using SCAR marker linked to R genes showed the presence of 4 resistant genes Co-2, Co-4, Co42, Co-6 in various resistant accessions. A maximum of 4 genes Co-2, Co-4, Co42, Co-6 were detected in 6 accessions. In the fungicides evaluation tests, trifloxystrobin 25 per cent+ tebuconazole 50 per cent (Nativo), tebuconazole (Folicur), and carbendazim 12 per cent + mancozeb 63 per cent (SAAF) were found highly effective whereas, among the botanicals aqueous extracts of M. azedarach was more effective against the C. lindemuthianum. While Trichoderma harzianum was more effective than Pseudomonas fluorescens and T. viride. The seed treatment with carbendazim (2.5 g/kg) followed by two foliar sprays of trifloxystrobin 25 per cent+ tebuconazole 50 per cent (Nativo), and T. harzianum at 45th and 60th day after sowing were found very effective against the disease in field trials and resulted in higher seed yield.
Open Access
VIRULENCE ANALYSIS OF Colletotrichum lindemuthianum AND ITS MANAGEMENT IN COMMON BEAN
(CSKHPKV, Palampur, 2017-07) SHARMA, NAIYA; Sharma, Dr. P.N.
ABSTRACT This study was aimed to determine pathogenic and molecular variability in Colletotrichum lindemuthianum, the casual organism of bean anthracnose; to evaluate bean germplasm to find out the sources of resistance against prevalent races; validation of R genes using molecular markers; and to identify suitable fungicide, biocontrol agents and botanicals for the management of disease. Virulence spectrum of 65 isolates determined on a set of 12 bean differential cultivars revealed the existence of 27 races in North Western Himalayas. Race 503 contained maximum number of 12 isolates from diverse geographic regions. Fifteen races viz; 5, 6, 7, 16, 18, 51, 87, 99, 145, 179, 211, 259, 337, 503 and 1395 were identified for the first time as none of them resembled with previously known races in Himachal Pradesh hence increasing the total number to 44 in the state. The isolates from J&K (3) and Uttrakhand (13) categorized into different races constitute their first record from these two states. The virulence analysis suggested 4 interaction types in accordance with the genetic origin of differential cultivars that were infected by a particular race. Twenty-one races having virulence for genotypes of both the gene pools showed Type IV interaction. Nine of 12 differential cultivars were found to be infected by one isolate suggesting 9 virulence factors among 65 isolates of C. lindemuthianum. RAPD data at 43 per cent similarity coefficient distinguished various isolates into 8 clusters, whereas, ISSR markers categorized test isolates into 5 clusters using 43 per cent similarity coefficient as a cutoff point. There was no congruence between the pathogenic variability and the molecular diversity data, showing no utility of such markers in differentiation of the physiological races whose identity is based on pathogenic behavior of the given isolate involving many pathogenicity factors. Out of 313 accessions, 45 were found resistant to race 0 (20), 17 (21), 503 (19) and 1395 (13), respectively. R-gene prediction analysis using SCAR marker linked to R genes showed the presence of 4 resistant genes Co-2, Co-4, Co42, Co-6 in various resistant accessions. A maximum of 4 genes Co-2, Co-4, Co42, Co-6 were detected in 6 accessions. In the fungicides evaluation tests, trifloxystrobin 25 per cent+ tebuconazole 50 per cent (Nativo), tebuconazole (Folicur), and carbendazim 12 per cent + mancozeb 63 per cent (SAAF) were found highly effective whereas, among the botanicals aqueous extracts of M. azedarach was more effective against the C. lindemuthianum. While Trichoderma harzianum was more effective than Pseudomonas fluorescens and T. viride. The seed treatment with carbendazim (2.5 g/kg) followed by two foliar sprays of trifloxystrobin 25 per cent+ tebuconazole 50 per cent (Nativo), and T. harzianum at 45th and 60th day after sowing were found very effective against the disease in field trials and resulted in higher seed yield.
Open Access
GENETICS AND MAPPING OF ANTHRACNOSE RESISTANCE GENE(s) IN COMMON BEAN LANDRACE KRC5
(CSKHPKV, Palampur, 2015-08-14) Katoch, Abhishek; Sharma, P. N.
ABSTRACT KRC5, a local landrace of common bean native to Kinnaur region of north western Himalayan state Himachal Pradesh is a well known for its effectiveness against eight pathogen races of C. lindemuthianum viz., 73, 83, 903, 591, 647, 775, 931 and 935. This study reports the identification, mapping and physical delimitation of the chromosomal location of a new anthracnose resistance gene Co-ind from landrace KRC5. The segregation analysis of an F2 progeny of a cross between a susceptible cv. ‘Jawala’ and resistant KRC5, an Andean genotype with three different races viz., 3, 537, 935 suggested that the resistance was conditioned by a single dominant gene. Bulk segregant analysis and recessive class analysis based strategy was used for the mapping of R gene. Preliminary mapping by linkage analysis of 159 F2/ RIL individuals with marker OPF6522 and OPR151136 localized the anthracnose resistance gene on chromosome 7. The R-locus was bracketed between the markers ScOPF6522 and OPR151136 located at a distance of 4.4 and 5 cM, respectively. In this region, six putatively expressed, LRR kinases and serine/threonine kinases (SRK) were short-listed as a potential candidate for the resistance gene identified from KRC5. Out of 178 accession phenotypically evaluated for resistance to race 3 of C. lindemuthianum, only five accessions viz., IC260292, IC265938, IC47839, EC400403 and EC325078 exhibited the presence of Co-ind. In R-gene postulation assay using SCAR markers SCO08, SZ20, SF10 and SAS13, bean accession IC262769 was identified as natural gene pyramid harboring four anthracnose resistance genes (Co-4, Co-42, Co-6 and Co-10) whereas 10 accessions viz., IC260336, IC258273, IC262748, IC47839, EC500328, EC4003433, EC325078 IC265940, IC202271 and IC265938 showed the presence of three resistance genes each. These multigenic accessions represent potential resistance sources and can be effectively utilized either directly as varieties after agronomic evaluation or can be involved in hybridization programmes to transfer resistance in susceptible genotypes after further validation.
Open Access
ANTI PLANT PATHOGENIC PROPERTIES OF HIGHER FUNGI ESPECIALLY WILD MUSHROOMS
(CSK Himachal Pradesh Krishi Vishavavidyalaya, Palampur, 2012) PANDEY, VIVEK KUMAR; Y.S. Paul
The present investigations on the anti plant pathogenic activity of wild mushrooms were undertaken to study characterization of fleshy fungi for anti plant pathogenic properties. Survey work was conducted in different forest areas of three districts of H.P. (Kangra, Mandi and Kullu). A total of 70 species of wild mushrooms belonging to 54 genera were collected and identified, out of which 40 species were cultured. Thirty two genera were recorded to be inedible, 14 as edible and four as poisonous. Boletus sp., Polyporus sp., Ganoderma lucidum and Tricholoma sp. were the most frequently distributed mushrooms. Antagonistic activities of 40 species of wild mushrooms against ten plant pathogenic fungi were tested by dual–culture technique. Most of the tested mushroom species markedly inhibited mycelial growth of all the phytopathogenic fungi. However, Cordyceps sp., Marasmius sp., Stropharia sp. and Russula sp. were found to have strong antagonistic activity. Culture filtrates of eleven wild mushrooms were found effective against test plant pathogenic fungi and nine were found effective against the bacterium Ralstonia solanacearum. Complete inhibition (100%) of Rhizoctonia solani and Sclerotinia sclerotiorum was observed by Russula sp., while maximum inhibition zone (30mm) was recorded by Stropharia sp. in Ralstonia solanacearum. Eleven culture filtrates and seventeen fruit body extracts of wild mushrooms were found to inhibit the conidial germination of Alternaria brassicae, Fusarium oxysporum, Colletotrichum capsici and Rhynchosporium oryzae. Minimum inhibition concentration of culture filtrates and fruit body extracts ranged between 50-80% and 25-80% concentration, however fruit body extract of Phellinus sp., Russula sp., and Amanita sp. completely (100%) inhibited the spore germination of Alternaria brassicae. Complete inhibition was also found in Rhynchosporium oryzae by Lactarius sp., Russula sp., Coprinus sp. and Cordyceps sp. The culture filtrates found effective under in vitro were also tested for host parasite interaction under pots conditions and found to reduce the disease incidence. Minimum disease incidence (26.66, 20.33 and 33%) was found in sheath and leaf blight of maize, stalk rot of cabbage and pea wilt by Russula sp. and Phellinus sp., respectively. In biochemical tests, total phenolics (1.60-22.70 mg/100ml) were the major antioxidant components found in the culture filtrates, followed by DPPH scavenging (2.50 -7.66) and hydroxyl radicals (2.59 - 5.2). Stropharia sp. contained the highest phenolic content (22.70 mg/100ml) while Cordyceps sp. contained highest concentration (7.66 and 5.52 mg/100ml) of DPPH scavenging and scavenging effect on hydroxyl radicals among the culture filtrates tested

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