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Open Access
Evaluation of Piriformospora indica against Piper yellow mottle virus in Black pepper
(Department of Plant Pathology, College of Agriculture, Vellayani, 2021) Lekshmi, R Krishnan; KAU; Joy, M
The study entitled “Evaluation of Piriformospora indica against Piper yellow mottle virus (PYMoV) in black pepper” was conducted in Department of Plant Pathology, College of Agriculture, Vellayani, Thiruvananthapuram during 2019-2021, to evaluate the efficiency of the root colonizing endophytic fungus P. indica in managing PYMoV in black pepper, and to elucidate the role of Reactive Oxygen Species and antioxidant enzymes in the tripartite interaction. A survey conducted in the instructional farm of College of Agriculture, Vellayani, to identify and collect PYMoV-infected black pepper plants showed that the surveyed varieties viz., Panniyur 1 to 7 and Karimunda were moderately to highly susceptible to PYMoV infection, with the disease incidence ranging from 10 to 91 per cent and disease severity as vulnerability index from 30 to 58. PYMoV-infected black pepper plants showed symptoms like chlorotic flecking, mottling, light necrosis, crinkling and distortion of leaves, reduction in leaf size and stunted growth. Presence of the virus in the samples was confirmed by polymerase chain reaction (PCR) with oligonucleotide primers specific to open reading frame III (ORF III) of PYMoV. P. indica culture obtained from Department of Plant Pathology was maintained in potato dextrose agar (PDA) and potato dextrose broth (PDB) media. P. indica was mass multiplied in coirpith-dried farm yard manure (FYM) mixture (1:1) amended with two per cent gram flour and was allowed to colonize on rooted cuttings of black pepper. P. indica colonization enhanced different biometric characters at different intervals, compared to the non-colonized control plants. P. indica colonization resulted in 25 per cent more shoot length, 23 per cent more root length, 35 per cent more number of leaves, 23 per cent more leaf area and 27 per cent more number of primary roots than the control plants at 150 days after co-cultivation (DAC). A pot culture experiment was laid out to evaluate the efficiency of prophylactic as well as curative application of P. indica, in managing PYMoV of black pepper. P. indica colonization significantly reduced the incidence and severity of PYMoD, upon pre- as well as post- inoculation of the virus, compared to the control plants. Plants colonized with P. indica and later inoculated with PYMoV as well as plants infected by PYMoV and later colonized by P. indica, both recovered the loss in biomass caused by the virus infection, compared to the non-colonized plants challenged by the virus. However, prophylactic treatment with P. indica resulted in better control of the disease compared to the curative treatment. Amplification of the virus by PCR also revealed that the virus titer was the least in P. indica-colonized plants that were post-inoculated with the virus after 30 DAC, followed by pre-inoculated plants that were later colonized with P. indica. The virus titer was the highest in the non-colonized plants inoculated with the virus. In order to evaluate the performance of P. indica in modulating the biochemical responses of black pepper plants against PYMoV infection, the activity of various enzymes which are known to play a role in the antioxidant defense systems of plants viz. catalase, peroxidase, glutathione reducatse, glutamate synthase, phosphatase and superoxide dismutase were assayed. A significant increase in the activity of these antioxidant enzymes was observed in the P. indica-colonized plants compared to the control plants, upon pre- as well as post- inoculation of PYMoV. Thus, the better performance of P. indica-colonized plants upon PYMoV infection can be correlated to the increased activities of antioxidant enzymes. PYMoV-infected plants showed increased accumulation of reactive oxygen species like superoxide anions and hydrogen peroxide, which cause oxidative stress in the plants. P. indica-colonization was able to mitigate the increase in ROS concentration caused due to PYMoV infection, thus reducing the symptoms. Thus the study reveals that, co-cultivation of rooted cuttings of black pepper (obtained from disease-free mother plants) with P. indica and keeping them under insectproof conditions for around 30 days (for successful establishment of the fungus inside the root cortex) and then planting them in the main field is a reliable and chemical-free method to control PYMoD in black pepper. Bio-protective action of P. indica might be attributed to the increased activity of antioxidant enzymes, which in turn leads to suppression in ROS accumulation in the colonized plants, upon being challenged by the virus.
Open Access
Varietal screening and management of anthracnose of black pepper using new generation fungicides
(Department of Plant Pathology, College of Agriculture, Vellayani, 2021) Athira, K; KAU; Heera, G
The study entitled “Varietal screening and management of anthracnose of black pepper using new generation fungicides” was conducted at Department of Plant Pathology, College of Agriculture, Vellayani during 2018 - 2020 with the objective to screen KAU varieties and most popular local cultivar Karimunda for resistance against anthracnose of black pepper caused by Colletotrichum gloeosporioides (Penz.) Penz. and Sacc. and evolve management strategy using new generation fungicides. As a part of the study, diseased samples were collected from three black pepper growing tracts of Kerala viz., Thiruvananthapuram, Wayanad and Idukki. Sample collections were made from two locations from Thiruvananthapuram (Kowdiar and Vellayani), Wayanad (Meenangadi and Ambalavayal) and Idukki (Myladumpara, Pampadumpara, Kattapana and Kambilikandam). Disease incidence and severity were assessed from the surveyed locations. The highest percentage disease index was observed in Myladumpara (50.28%) followed by Kattapana (48.62%). Weather parameter viz., temperature, relative humidity and rainfall were recorded during the survey period. The weather parameters viz., low temperature, high relative humidity and heavy rainfall favoured the incidence of anthracnose. The symptoms of the anthracnose appeared as small necrotic spots with a yellow halo on the leaf lamina. Several lesions coalesce together resulted in leaf blight and defoliation. In Pampadumpara, spike infection was also observed along with leaf spot. Colletotrichum cultures were isolated from the diseased sample by tissue isolation technique and eight pure cultures of Colletotrichum sp. (C1 to C8) were obtained. The pathogenicity of the eight isolates of Colletotrichum sp. from different locations were proved by Koch postulates. The morphological characters of the eight different isolates were studied in potato dextrose agar (PDA) medium. The isolated cultures of Colletotrichum sp. produced whitish with yellowish orange centre to light pink, off white to greyish coloured colony having fluffy, cottony to sparse mycelial growth with regular margins. Days taken to grow the entire petridish ranged from 7.25 to 9.75 days. The mycelium of the fungus was hyaline and septate, and its width ranged from 2.21 - 3.45 μm. The septal distance of the different Colletotrichum isolates ranged between 8.50 - 21.23 μm. The conidia were single celled with an oil globule at the centre. The conidial shape was either cylindrical, oblong or dumbbell. The conidial and appressorial size varied from 9.4 - 12.1 μm x 3.6 - 4.6 μm and 8.5 – 11.2 μm x 3.5 – 4.3 μm respectively. The isolates were identified as Colletotrichum gloeosporioides. The pathogenic variability of the eight C. gloeosporioides isolates were assessed on three black pepper varieties viz., Panniyur 1, Panniyur 3 and Karimunda by virulence rating. The isolate C7 was identified as the most virulent isolate which produced lesion size of 1.92 cm, 2.40 cm, and 3.22 cm on Panniyur 1, Panniyur 3 and Karimunda respectively at 5 days after inoculation (DAI). The isolate C7 produced symptoms within two days after artificial inoculation in the three varieties tested with a higher rate of lesion development of 0.40 (Panniyur 1), 0.49 (Panniyur 3) and 0.66 (Karimunda) cm day-1. KAU varieties (Panniyur 1 to 8) and local cultivar Karimunda were screened against the most virulent isolate of C. gloeosporioides. Among the varieties screened, Panniyur 4 was found to be highly susceptible with highest PDI of 51.43 (7 DAI), whereas Panniyur 2 had the lowest PDI of 14.28 (7DAI) followed by Panniyur 8 with PDI 20.00 % (7DAI) and were found to be tolerant to anthracnose infection. Panniyur 1, Panniyur 7 and Panniyur 5 were also found to be moderately susceptible. The pathogen produced symptoms in susceptible varieties within 2 DAI, whereas the tolerant varieties took 3-4 days to initiate the infection. In vitro screening of new generation fungicides revealed that kresoxim methyl of strobilurin and tebuconazole of triazole were the most effective in inhibiting mycelial growth of C. gloeosporioides (80.37% and cent percent respectively). The combination fungicide carbendazim 12% + mancozeb 63 % completely inhibited the mycelial growth at 25, 50 and 100 ppm. The combination fungicides azoxystrobin 11% + tebuconazole 18.3% SC and trifloxystrobin 25% + tebuconazole 55% WP were also effective against the pathogen at 100 ppm. The contact fungicide copper oxychloride was ineffective against the pathogen @ 10, 25, 50 and 100 ppm. The present study revealed the use of tolerant varieties along with need based application of new generation fungicides to keep the destructive disease under control. The future line of work should include screening of more black pepper varieties under field condition to assess their reaction to anthracnose, elucidation of the factors governing resistance to the disease and the efficacy of new generation fungicides under field condition.
Open Access
Eco-friendly management of Fusarium rot in cardamom and its impact on soil health and plant defense mechanism
(Department of Plant Pathology, College of Agriculture, Vellayani, 2021) Veni Krishna, K C; KAU; Dhanya, M K
Open Access
Etiology and characterization of diseases of Anthurium (Anthurium andraeanum L.) in Kerala
(Department of Plant Pathology, College of Agriculture, Vellanikkara, 2021) Nitha, Rafi; KAU; Anita Cherian, K
Anthurium (Anthurium andraeanum L.), a native of Tropical America is a perennial herbaceous plant commercially grown for its attractive coloured spathe and green shiny foliage. These have great economic value in the global flower trade that led to the budding of many urban and rural anthurium entrepreneurs in the countryside, particularly in the state of Kerala. During the recent past, the cultivation of this high valued cut flower crop is challenged by many biotic factors and the changes in climate scenario occurred recently led to the emergence of various new pests and diseases. Hence the present study was undertaken to identify and document the diseases affecting anthurium grown in the state of Kerala and the characterization of associated pathogens. Purposive sampling surveys were conducted in the anthurium cultivated locations of six districts viz. Thrissur, Ernakulam, Palakkad, Malappuram, Kozhikode and Wayanad during the period from October, 2020 to July, 2021. The incidence and severity of various symptoms were assessed and collected the samples for further studies. Twelve leaf spots (KMALS, PBALS, VFNLS, VCNLS1, VCNLS2, IJKLS, KKYLS, OKMLS, OLRLS, TLRLS, TBMLS and NLBLS) two leaf blights (VFNLB and OLKLB), one root rot (VCNRR), one wilt (MNTLW) four inflorescence rots (MNTSR, CKDSR, PNMSR and ALVSR) and one mosaic symptom (VCNML) were collected during the survey. Among the leaf spots, PBALS recorded the highest per cent disease severity (PDS) of 58.23 followed by NLBLS (46.66 %) and ALVSR recorded the highest severity of 32 per cent among inflorescence rots. Isolation and pathogenicity studies of the associated pathogens yielded 18 fungal and two bacterial isolates. Symptomatology of these diseases were studied both under natural and artificial conditions. The fungal pathogens were characterized and identified by studying cultural and morphological characteristics. Based on these characters, leaf spot pathogens were identified as Colletotrichum gloeosporioides, Lasiodiplodia theobromae, Phoma sp., Phomopsis sp., Corynespora sp., Pestalotiopsis sp. and those causing inflorescence rot as Colletotrichum gloeosporioides and Phomopsis sp. and root rot pathogen as Phytopythium sp. and wilt pathogen as Fusarium sp. The bacterial isolates causing leaf blight were identified as Xanthomonas sp. based on cultural, morphological and biochemical characteristics. The viral like symptoms subjected to electron microscopy revealed the absence of any virus particles. PCR amplification of Internal Transcribed Spacer (ITS) region of the fungal genome followed by sequencing and in silico analysis confirmed that the pathogen associated with TBMLS as C. queenslandicum, VCNRR as Phytopythium vexans, IJKLS as Diaporthe phaseolarum, KKYLS and OKMLS as Phomopsis heveicola, NLBLS as Lasiodiplodia theobromae, TLRLS as Pseudopestalotiopsis thea, VCNLS2 and OLRLS as Corynespora cassiicola, MNTLW as Fusarium fujikuroi. The pathogens associated with KMALS, PBALS, VFNLS, MNTSR, PNMSR and ALVSR belong to Colletotrichum gloeosporioides species complex. The bacterial isolates (VFNLB and OLKLB) were confirmed as Xanthomonas axonopodis through the amplification of 16S rRNA region of the genome followed by sequencing and in silico analysis. An in vitro experiment was conducted to study the efficacy of fungicides and biocontrol agents/bioformulations against major fungal pathogens. Fungicides viz. propineb, difenoconazole, carbendazim 12 % + mancozeb 64 %, cymoxanil 8% + mancozeb 64 % and Bordeaux mixture were highly effective against L. theobromae and Colletotrichum sp. (PBALS) as it resulted in cent per cent inhibition of these two pathogens at all the three doses. In the case of P. vexans, fungicides viz. propineb, azoxystrobin, carbendazim 12 % + mancozeb 64 %, cymoxanil 8% + mancozeb 64 % and Bordeaux mixture were found to be effective even at a lower dose. Among the three chemicals tested against Xanthomonas axonopodis, maximum inhibition was recorded in the case of streptocycline. Dual culture and filter paper disc method revealed that the biocontrol agents/formulations viz. Trichoderma sp., PGPR-II and PGPM were effective against all the tested fungal and bacterial pathogens. Pseudomonas fluorescens was found effective against three fungal and the bacterial pathogen.
Open Access
Host range studies and management of anthracnose of nutmeg caused by colletotrichum spp.
(Department of Plant Pathology, College of Agriculture, Vellayani, 2020) Bommana, Divya; KAU; Heera, G
The study entitled “Host range studies and management of anthracnose of nutmeg caused by Colletotrichum spp .” was conducted at Department of Plant Pathology, College of Agriculture, Vellayani during 2018-2020 with the objectives to characterize the causal agent, study the host range of Colletotrichum spp. causing anthracnose and to develop effective management strategy to control the disease by using new generation fungicides. As a part of the study, anthracnose infected samples were collected from four nutmeg growing districts of Kerala viz., Thiruvananthapuram, Kottayam, Ernakulam and Idukki. For the infected sample collections, three locations from Thiruvananthapuram (Vellayani, Karamana and Palode), two locations from Kottayam (Kumarakom and Vaikom), six locations from Idukki (Myladumpara, Pampadumpara, Adimali, Kambilikandam, Panickankudy and Kattapana) and one location from Ernakulam (Kadungalloor) were surveyed. Disease incidence and severity were assessed from the surveyed locations. The highest disease incidence and severity were observed in Kadungalloor (DI - 90 % and PDI - 56.40 respectively) followed by Kumarakom (DI - 80 % and PDI - 41.33 respectively) and the lowest disease incidence and severity in Myladumpara (DI - 20 % and PDI - 15.53 respectively). The symptoms of the anthracnose on nutmeg appeared as small necrotic spots with a prominent yellow halo on the leaf lamina. Several lesions coalesced together resulted in leaf blight, shot hole and defoliation. In Kambilikandam and Panickankudy. fruit rot was also observed along with leaf spot. The cultures of Colletotrichum spp. were isolated from the infected samples from different locations. Eighteen pure cultures of Colletotrichum sp. (C1 to C18) were obtained. Seven isolates of Colletotrichum sp. were selected for further studies based on the days taken for symptom development and rate of lesion development. The pathogenicity of the seven isolates of Colletotrichum sp. from different locations were proved by Koch postulates. The morphological and culture characters of the seven different isolates were studied in potato dextrose agar (PDA) medium. The isolated cultures of Colletotrichum sp. produced whitish to greyish radiating mycelium; later turning to off white to pink coloured fluffy to sparse mycelium with regular margins. Days taken to grow the entire Petri dish ranged from 7 to 10. The mycelium of the fungus was hyaline and septate; and its width ranged from 0.46 μm to 2.48 μm. The conidia were single celled with an oil globule at the centre and wereoblong or dumbbell shaped. The conidial size varied from 7.87 to 19.97 μm x 3.26 to 5.68 μm. The isolates were morphologically identified as C. gloeosporioides. The pathogenic variability of the seven isolates of C. gloeosporioides was assessed on detached nutmeg twigs by virulence rating. The isolate C4 was identified as the most virulent isolate which produced lesion size of 11.89 cm and 16.81cm at 7 DAI and 9 DAI respectively. The isolate C4 produced symptoms within two days after artificial inoculation and had a higher rate of lesion development of 4.12 cm day -1 . The other isolates took 3 to 4 days for symptom appearance on artificial inoculation of the pathogen. Host range of the most virulent isolate of C. gloeosporioides (C4) obtained from nutmeg was studied in perennial tree spices viz., clove, cinnamon, all spice, betel vine, black pepper and coconut. C. gloeosporioides isolate of nutmeg is capable of infecting the above- mentioned host plants. The isolate produced symptoms in all the hosts within 2 to 4 DAI and the symptoms developed varied from brown lesions, brown lesions with a shot hole to necrotic spots with prominent yellow halo. The maximum lesion size of 2.43 cm was observed in clove and minimum lesion size of 1.31 cm in all spice. In vitro screening of new generation fungicides revealed that triazole group fungicide propiconazole 25EC at 100 ppm and combination fungicides, carbendazim 12% + mancozeb 63% at 25 ppm; and Trifloxystrobin 25% + Tebuconazole 55% WP at 100 ppm concentration were the most effective in completely inhibiting the mycelial growth of the pathogen. The present study revealed the wide host range of the C. gloeosporioides isolate of nutmeg and also the effectiveness of new generation fungicides in managing the pathogen. The future line of work should include molecular variability between various isolates, cross infectivity among the isolates in other perennial hosts, and the efficacy of new generation fungicides under field condition.
Open Access
Varietal screening and management of anthracnose of black pepper using new generation fungicides
(Department of Plant Pathology, College of Agriculture, Vellayani, 2020) Athira, K; KAU; Heera, G
The study entitled “Varietal screening and management of anthracnose of black pepper using new generation fungicides” was conducted at Department of Plant Pathology, College of Agriculture, Vellayani during 2018 - 2020 with the objective to screen KAU varieties and most popular local cultivar Karimunda for resistance against anthracnose of black pepper caused by Colletotrichum gloeosporioides (Penz.) Penz. and Sacc. and evolve management strategy using new generation fungicides. As a part of the study, diseased samples were collected from three black pepper growing tracts of Kerala viz., Thiruvananthapuram, Wayanad and Idukki. Sample collections were made from two locations from Thiruvananthapuram (Kowdiar and Vellayani), Wayanad (Meenangadi and Ambalavayal) and Idukki (Myladumpara, Pampadumpara, Kattapana and Kambilikandam). Disease incidence and severity were assessed from the surveyed locations. The highest percentage disease index was observed in Myladumpara (50.28%) followed by Kattapana (48.62%). Weather parameter viz., temperature, relative humidity and rainfall were recorded during the survey period. The weather parameters viz., low temperature, high relative humidity and heavy rainfall favoured the incidence of anthracnose. The symptoms of the anthracnose appeared as small necrotic spots with a yellow halo on the leaf lamina. Several lesions coalesce together resulted in leaf blight and defoliation. In Pampadumpara, spike infection was also observed along with leaf spot. Colletotrichum cultures were isolated from the diseased sample by tissue isolation technique and eight pure cultures of Colletotrichum sp. (C1 to C8) were obtained. The pathogenicity of the eight isolates of Colletotrichum sp. from different locations were proved by Koch postulates. The morphological characters of the eight different isolates were studied in potato dextrose agar (PDA) medium. The isolated cultures of Colletotrichum sp. produced whitish with yellowish orange centre to light pink, off white to greyish coloured colony having fluffy, cottony to sparse mycelial growth with regular margins. Days taken to grow the entire petridish ranged from 7.25 to 9.75 days. The mycelium of the fungus was hyaline and septate, and its width ranged from 2.21 - 3.45 μm. The septal distance of the different Colletotrichum isolates ranged between 8.50 - 21.23 μm. The conidia were single celled with an oil globule at the centre. The conidial shape was either cylindrical, oblong or dumbbell. The conidial and appressorial size varied from 9.4 - 12.1 μm x 3.6 - 4.6 μm and 8.5 – 11.2 μm x 3.5 – 4.3 μm respectively. The isolates were identified as Colletotrichum gloeosporioides. The pathogenic variability of the eight C. gloeosporioides isolates were assessed on three black pepper varieties viz., Panniyur 1, Panniyur 3 and Karimunda by virulence rating. The isolate C7 was identified as the most virulent isolate which produced lesion size of 1.92 cm, 2.40 cm, and 3.22 cm on Panniyur 1, Panniyur 3 and Karimunda respectively at 5 days after inoculation (DAI). The isolate C7 produced symptoms within two days after artificial inoculation in the three varieties tested with a higher rate of lesion development of 0.40 (Panniyur 1), 0.49 (Panniyur 3) and 0.66 (Karimunda) cm day-1. KAU varieties (Panniyur 1 to 8) and local cultivar Karimunda were screened against the most virulent isolate of C. gloeosporioides. Among the varieties screened, Panniyur 4 was found to be highly susceptible with highest PDI of 51.43 (7 DAI), whereas Panniyur 2 had the lowest PDI of 14.28 (7DAI) followed by Panniyur 8 with PDI 20.00 % (7DAI) and were found to be tolerant to anthracnose infection. Panniyur 1, Panniyur 7 and Panniyur 5 were also found to be moderately susceptible. The pathogen produced symptoms in susceptible varieties within 2 DAI, whereas the tolerant varieties took 3-4 days to initiate the infection. In vitro screening of new generation fungicides revealed that kresoxim methyl of strobilurin and tebuconazole of triazole were the most effective in inhibiting mycelial growth of C. gloeosporioides (80.37% and cent percent respectively). The combination fungicide carbendazim 12% + mancozeb 63 % completely inhibited the mycelial growth at 25, 50 and 100 ppm. The combination fungicides azoxystrobin 11% + tebuconazole 18.3% SC and trifloxystrobin 25% + tebuconazole 55% WP were also effective against the pathogen at 100 ppm. The contact fungicide copper oxychloride was ineffective against the pathogen @ 10, 25, 50 and 100 ppm. The present study revealed the use of tolerant varieties along with need based application of new generation fungicides to keep the destructive disease under control. The future line of work should include screening of more black pepper varieties under field condition to assess their reaction to anthracnose, elucidation of the factors governing resistance to the disease and the efficacy of new generation fungicides under field condition.
Open Access
Eco-friendly management of Fusarium rot in cardamom and its impact on soil health and plant defence mechanism
(Department of Plant Pathology, College of Agriculture, Vellayani, 2020) Veni Krishna, K C; KAU; Dhanya, M K
The study entitled ‘Eco-friendly management of Fusarium rot in cardamom and its impact on soil health and plant defense mechanism’ was conducted at College of Agriculture, Vellayani and Cardamom Research Station Pampadumpara during 2018- 2020 with the objective to assess Fusarium rot severity and pathogen variability in Idukki district, develop an effective ecofriendly management practices for the disease and study the impact of the practices on the soil and plant health. Six isolates of Fusarium oxysporum Schlecht were obtained, three each from infected pseudostem and roots collected from Kattapana and Nedumkandam blocks of Idukki district. The isolate, Fp1 from pseudostem (Pampadumpara panchayath) was identified as most virulent based on virulence rating (viz., days taken for symptom development, lesion length, mycelial growth), and was used for further studies. Inoculation of the root isolate (Fr1) also produced typical lesion on pseudostem indicating that the, fungus can also attack pseudostem besides the roots. But the root isolate took more days (40 days) for symptom development compared to pseudostem isolate (29 days). The morphological characters including the colour, colony characters, hyphal and conidial characters as well as rate of growth in Petri dish confirmed the fungus as F. oxysporum. In vitro studies revealed that the isolates from pseudostem and roots showed significant variation in their morphological, cultural and pathological characters. A pot culture experiment was conducted to assess the efficacy of selected bioagents (individually and in combination) for the management of the disease at CRS Pampadumpara in CRD using nine treatments with three replications. Pathogen inoculum (150 gm/10 kg soil) multiplied in sand-maize flour medium was standardised as the inoculum level enough for cent per infection on the pseudostem resulting in complete crop loss. Soil application of vermiculite based AMF inoculum (20 g) with @ 2% Pseudomonas fluorescens (1 L/10 kg soil) per 10 kg soil at the time of planting along with 2% P fluorescens spray @ 0.5 L/ plant at monthly interval for three times resulted in effective disease management (disease incidence: 40% and disease severity: 24.26%) compared to the inoculated control (disease incidence: 100% and disease severity: 69.38 %). Studies on the population dynamics of the pathogen and the biocontrol agents at periodical interval upto three months revealed significant reduction in the pathogen antagonist ratio and enhancement in AMF colonization of the treatment plants. Among the best treatment the combination of AMF and P. fluorescens resulted in good biometric characters of the treatment plants (plant height: 96.50 cm ,leaf length: 62.50 cm and number of leaves: 15.75) compared to control (plant height: 41 cm ,leaf length: 30 cm and number of leaves: 7.50) through enhancement of soil nutrients (P, K, Ca and Mg) and plant nutrient status (K, Mg, S and B) compared to control plants. Laboratory studies also revealed the induction of defense related enzymes (phenol, ortho dihydroxy phenol, peroxidase, polyphenol oxidase, and β 1,3 glucanase) in high concentration as a response to the application of above treatments. Thus, the present study revealed that Fusarium rot of cardamom a devastating disease can be effectively managed by soil application of vermiculate based AMF inoculum (20 g) with 2 per cent P. fluorescens (1L/10 kg soil) per 10 kg soil at the time of planting along with 2 per cent P. fluorescens spray at 0.5 L per plant at monthly interval for three times; this treatment also resulted in good vegetative growth of cardamom plants therefore this can be used as an ecofriendly management strategy for the production of good quality cardamom.
Open Access
Strain improvement of Trichoderma spp. by protoplast fusion
(Department of Plant Patholgy, College of Agriculture, Vellayani, 2020) Anit, Cyriac; KAU; Sible George, Varghese
A study on “Strain improvement of Trichoderma spp. by protoplast fusion” was conducted at Department of Plant Pathology, College of Agriculture, Vellayani during the year 2018-2020, with the objective of improving the screened strains of Trichoderma spp. by protoplast fusion for spp. by protoplast fusion for increasing the antagonistic ability and related traits against soil borne pathogens. A survey was conducted in five agro-climatic zones of Kerala viz., Northern Zone, Central Zone, High Range Zone, Problem Area Zone and Southern Zone for collection of soil samples especially from forest soils. The collected soil samples were assessed for the population of Trichoderma spp. A total of 31 Trichoderma spp. isolates were obtained from the soil samples collected from five agro-climatic zones. Majority of the isolates were obtained from soils with pH of 6 to 7. Isolate TRMW2, TREN1, TREZ1, TREZ2, TREZ3, TRRN1, TRRN2, TRKR1, TRPN3, TRPN7, TRPN10 and TRPN18 exhibited full growth at four days after inoculation (DAI). The isolated Trichoderma spp. differed in growth rate and colony characters like colour of mycelium, texture of colony and sporulation pattern. Isolates of Trichoderma spp. from different zones exhibited in vitro inhibition against soil borne pathogens such as Pythium aphanidermatum and Rhizoctonia solani. Majority of the isolates displayed high inhibition per cent compared to KAU strain of Trichoderma sp. TRRN1, TRRN2, TRPN3, TRPN7, TRPN11, TRPN15 and TRKR2 isolates exhibited complete inhibition of P. aphanidermatum in dual culture experiment; whereas TREN1, TRMW2, TREZ1, TREZ2, TRKM1, TRPN7, TRPN9, TRPN14, TRPN15, TRPN17, TRPN18 and TRKR2 isolates exhibited complete inhibition of R. solani. Trichoderma isolates such as TRPN7, TRPN15 and TRKR2 exhibited complete inhibition against both the pathogens. Antagonistic properties viz., antibiosis, lysis and overgrowth of Trichoderma isolates against P. aphanidermatum and R. solani were observed. During the antagonist-pathogen interaction, isolates TRSN1, TRSN2, TRPN10, TRPN14, TRPN15, TRPN17 and TRPN18 exhibited high levels of antibiosis. Most of the isolates caused lysis of mycelium of the pathogens which resulted in formation of clear zones in dual culture. Overgrowth of the antagonist was another prominent antagonistic property observed in the majority of the isolates. Based on the antagonistic properties, the Trichoderma isolates viz., TRSN1, TRMW2 and TRPN14 were selected for the protoplast fusion. During the protoplast isolation, the maximum number of protoplasts was obtained after 2 h of incubation of mycelia of parental isolates with the lytic enzyme. Protoplast fusion was carried out between the selected isolates (TRSN1 x TRPN14, TRSN1 x TRMW2, and TRPN14 x TRMW2) in the presence of poly ethylene glycol (PEG 6000). Three protoplast fusants were selected using carbendazim-amended PDA medium. The protoplast fusants displayed fast growth on PDA medium and completely covered the Petri dish at 5th of growth. The colony characters of fusants varied from light to dark green mycelium with fluffy growth and scattered to circular green heavy sporulation. In vitro screening of protoplast fusants against P. aphanidermatum and R. solani revealed that highest inhibition against P. aphanidermatum was observed with fusant 3 (84.4%) followed by fusant 2 (74.44%). Highest inhibition against R. solani was observed with fusant 2 (100%) followed by fusant 3 (70.30%). Antagonistic properties viz., antibiosis, lysis and overgrowth were observed in the protoplast fusants. Among the three protoplast fusants, fusant 1 exhibited all the antagonistic properties against both the pathogens with heavy sporulation. Thus, the present study has thrown light in understanding the potential of protoplast fusion in evolving improved strains of Trichoderma spp. Protoplast fusion enhanced sporulation in fusants compared to the parents. Further studies need to be conducted for the biochemical and molecular characterisation of parental isolates and fusants. The parents and protoplast fusants also have to be evaluated for their in vivo efficacy against soil borne pathogens in major crops of Kerala.
Open Access
Characterization of fungal pathogen associated with leaf rot disease of coconut (Cocos nucifere L.) and In Vitro evaluation of phylloplane microflora as biocontrol agents
(Department of Plant Pathology, College of Agriculture, Vellayani, 2020) Deena, Sebastian; KAU; Radhakrishnan, N V
Leaf rot disease (LRD) is a major foliar disease affecting coconut plantations of Southern Kerala especially in root (wilt) affected areas. In this context, the study entitled ‘Characterization of fungal pathogen associated with leaf rot disease of coconut (Cocos nucifera L.) and in vitro evaluation of phylloplane microflora as biocontrol agents’ was conducted in the Department of Plant Pathology, College of Agriculture, Vellayani during the year 2018-2020, with the objective to identify and characterize the major fungal pathogens associated with the LRD of coconut and in vitro evaluation of phylloplane microflora of coconut against the pathogens. The isolation of LRD pathogens was carried out from six taluks of Thiruvananthpuram district such as Thiruvananthapuram, Neyyattinkara, Nedumangad, Chirayinkeezhu, Kattakada and Varkala. Three locations were selected from each taluk and a total of eighteen samples were collected during the study. The results revealed that the disease in Thiruvananthapuram district was caused by a spectrum of pathogens such as Colletotrichum gloeosporioides, Fusarium spp., Gliocladium sp., and Scytalidium sp. The LRD was caused either by a single pathogen or by combinations of pathogens. C. gloeosporioides and Fusarium spp. were found as the major pathogens of LRD based on the frequency of isolation. Each and every isolate of the same pathogen differed from one another in cultural characters and virulence. All the pathogens produced water soaked brown lesion on artificial inoculation on detached spindle leaves; though the days taken for symptom initiation and size of the lesion developed varied. The isolate C3 (Isolate from Anayara, Thiruvananthapuram taluk) was found to be more virulent among the C. gloeosporioides isolates; and among the Fusarium spp. isolates, the isolate F5 (Isolate from Alamkode, Chirayinkeezhu taluk) was found to be more virulent. By observing the spore characters of the isolates, it was found that the spore size and pigmentation of the culture haven’t any significance to the virulence of the pathogen. Dual inoculation of the major pathogens on detached spindle leaves caused severe incidence of the disease compared to the individual inoculation of the pathogens. This result indicated that the LRD caused by fungal complex is more severe than that caused by individual fungal isolates. There are phylloplane fungi existing on healthy leaves of the infected palm with enough inhibition potential to LRD. The phylloplane fungal isolate PF5 showed more per cent inhibition to mycelial growth of C. gloeosporioides (54.44%) followed by the isolate PF4 (43.33%); and the isolate PF4 showed more inhibition to Fusarium sp. (64.44%) followed by the isolate PF5 (45.55%) in the dual culture assay. The detached spindle leaf assay also supported the same fact that the isolate PF5 was observed to be more suppressive to the disease caused by C. gloeosporioides (28.77%) and the isolate PF4 was reported to have more suppression to the disease caused by Fusarium sp. (34.56%). These pre-treatment effects are more promising than Pseudomonas fluorescens PN026, but inferior to copper oxy chloride (0.2%). Thus, the present study revealed that the LRD of coconut in Thiruvananthapuram district is caused by a combination of pathogenic fungi viz., C. gloeosporioides, Fusarium spp., Gliocladium sp., and Scytalidium sp. Prophylactic application of the phylloplane fungal isolates PF4 and PF5 could reduce the LRD severity in vitro to a promising level and these isolate can be further tested for in vivo biocontrol potential before going for the development of a formulated product.

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