General Part-1

Technology Code:-	:	201713439520659
Organization Details...
Subject Matter Division	:	{{smdOb.smdName}}
Organization Name	:	{{orgOb.orgName}} ,{{orgOb.City}}
AICRP name if any (AICRP)	:	All Not Applicable {{aicrpOb.aicrpName}}
Division name if any	:	Crop Protection
Details of Inventors..
Principal Inventor	:	A. Ishwara Bhat
Principal Inventor Designation:	:	Principal Scientist
Principal Inventor Email	:	ishwarabhat.a@icar.gov.in
Principal Inventor Address	:	Principal Scientist ICAR-Indian Institute of Spices Research Marikunnu Post Kozhikode 673012 Kerala
Co-Inventor Name	:	Shina Sasi
Co-Inventor Email	:	shinasasi88@gmail.com
Technology Name	:	Elimination of Piper yellow mottle virus (PYMoV) from virus-infected black pepper and production of virus-free plants through somatic embryogenesis and meristem-tip culture
Technology Details..
Major resource	:	All Not Applicable {{majorOb.majreName}}
Minor Subject Classification	:	All Not Applicable {{minorOb.minorName}}
Minor Subject Sub Classification	:	All Not Applicable {{minorOb.minorsubName}}
Technology Group	:	All Not Applicable {{techgroup.tecg_group_name}}
Technology Related To	:	All Not Applicable {{techrela.tr_name}}
Complete Details of Technology:	:
Piper yellow mottle virus (PYMoV) is a pararetrovirus associated with stunt disease in black pepper. As the primary spread of the virus occurs through vegetative propagation, the elimination of the virus from the infected plant and the production of virus-free plants is necessary to check the spread of the virus. Currently, no methods are available to eliminate the virus from the PYMoV-infected black pepper plants. Keeping this in view, in this study, somatic embryogenesis and meristem-tip culture alone and in combination with the antiviral agent ribavirin were attempted for the elimination of PYMoV from infected black pepper explants. Cyclic somatic embryo obtained from the micropylar region of mature seeds from PYMoV-infected black pepper plants of six varieties were regenerated and hardened in the greenhouse. Testing of somatic embryo-derived plants by PCR showed virus elimination in 55−100% of the plants in different varieties of black pepper. Increased PYMoV elimination was obtained when cyclic somatic embryos were pre-treated with ribavirin before regeneration. A protocol for meristem-tip culture of black pepper plants was developed that consisted of excising the meristem from PYMoV-infected black pepper plants and inoculating in the regeneration medium containing antibiotics (to remove endophytic bacterial contamination), followed by rooting and hardening of the plants. Testing of meristem-derived plants showed PYMoV elimination in 84% of the plants. Further elimination of the virus was obtained when the meristem-tip culture was combined with ribavirin treatment.
Brief Description of Technology Including Salient Features:
The method involved inoculation of the embryo along with the micropylar region (obtained from matured seeds collected from PYMoV-infected black pepper plants) on SH30 medium (SH medium containing 3% sucrose) and incubated at 28 °C in dark condition for the induction of primary somatic embryogenesis. The primary somatic embryo along with the whole explant was inoculated in SH10 for the induction of secondary somatic embryogenesis and cyclic somatic embryogenesis. The cyclic somatic embryo were treated with 20 mg/l of ribavirin and regenerated to produce plantlets in SH35 liquid medium (SH medium containing 3.5% sucrose) with shaking at 110 rpm for 30 days. Well-differentiated plantlets were transferred to a woody plant medium (WPM) and rooted plantlets were hardened in the greenhouse. Somatic embryo-derived plants were tested for the presence/absence of PYMoV by PCR. The protocol for meristem-tip culture involved the collection of shoot tips of PYMoV-infected black pepper plants followed by treatment with water containing 1% Tween-20 for 10 min and washed under running tap water for 10 min. Explants were surface sterilized using 0.1% mercuric chloride for 10 min and meristems of approximately 2-3 mm were excised from the shoot tips under laminar air flow using a clean, sterile scalpel and were inoculated into WPM medium containing 3 mg/l benzyl adenine (BA) + 1 mg/l kinetin (KN), +1% tetracycline +1% spectromycin and 10 mg/l ribavirin for a period of 30 days, with two subculturing in the same medium at an interval of 15 days. The rooted plantlets were hardened in the greenhouse and tested for the presence/absence of PYMoV by PCR. The virus-free plants obtained were then used as mother plants for further multiplication under insect-proof conditions and used for planting in the field.
Benefits/Utility	:
The virus infection can cause a yield reduction of up to 82% depending on the severity of the disease. The protocol developed by us can be used for the production of PYMoV-free plants of black pepper. Planting virus-free plants in the field ensures healthy crops without disease which contributes to an increase in productivity.
Precaution With The Technology	:	The somatic embryos and meristem cultures are prone to contamination under in vitro conditions. Hence regular checking of somatic embryos and meristem cultures and discarding the contaminated ones are needed. The virus-free plants from the above methods should be propagated under an insect-proof nursery to avoid virus spread through insects such as mealybugs.
How To Use	:
The virus-free plants obtained were used as mother plants in the insect-proof nursery for further propagation through the serpentine method. The virus-free plants obtained from the nursery are then used for planting in the main field.
Impact, If Adopted	:
The technology ensures the production of PYMoV-free plants from PYMoV-infected plants of black pepper. The planting of virus-free plants in the field ensures healthy crops without disease which contributes to an increase in productivity.
Social Impact	:
The virus infection can cause a yield reduction of up to 82% depending on the severity of the disease. The protocol developed can be used for the production of PYMoV-free plants of black pepper. Planting virus-free plants in the field ensures healthy crops without disease which contributes to an increase in productivity.
TargetUsers/Stake holders	:	Tissue culture laboratories involved in the multiplication of black pepper, farmers, breeders.
Technology Contact..
Name	:	Director
Email	:	director@spices.res.in,director.spices@icar.gov.in
Phone Number	:	0495-2730294
Fax Number	:	0495-2731187
Address	:	ICAR-Indian Institute of Spices Research,Marikunnu P.O.,Calicut-673012
Keyword for Technology	:	Somatic embryogenesis, meristem tip culture, virus-free plants, tissue culture, black pepper, piper yellow mottle virus, detection

Technology Development Details Part-2

Project Details (Through which technology was developed)	:	PhD research project
Time of Initiation Technology Development	:	1-2013
Time of Completion Technology Development	:	2-2016
Technology Validated by	:	Within ICAR
Technology Validation Details..
Subject Matter Division	:	{{smdOb.smdName}}
Organization Name(if within ICAR)	:	ICAR-Indian Institute of Spices Research,Calicut
Organization Name(if outside ICAR,Please enter)	:
Year of Validation(YYYY)	:	2-2015
Year of Release/Adoption(YYYY)	:	3-2016
Country	:	India

Applies To(Regional Differentiation)Inform Part-3

Location...
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Sub zone(As per the planning commission)	:	All Not Applicable {{zonesub.agroName}}, {{zonesub.Region}}
AgroEcological Zone(NBSS & LUP)	:	All Not Applicable {{agrozone.nbssaerName}}
AgroEcological Sub Zone(NBSS & LUP)	:	All Not Applicable {{agrosubzone.nbssaesrName}}
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District Name	:	All Not Applicable {{dist.distName}}
Soil Type/Resource Type..
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Soil Sub Order	:	All Not Applicable {{soilsuborder.soilsubName}}
Soil great group	:	All Not Applicable {{soilgreat.soilgreatName}}
Soil great sub group	:	All Not Applicable {{soilgreatsub.soilgreatsubName}}
Commodity Details..
Commodity	:	All Not Applicable {{commodity.commodityName}}
Commodity Type	:	All Not Applicable {{commoditytype.commoditytypeName}}
Commodity Name	:	All Not Applicable {{commodityname.commodityName}}

Publication Related To Technology Part-4

Publication Related To Technology Part-4

Research Paper information..

1. Sasi S and Bhat A I (2018 ). In vitro elimination of Piper yellow mottle virus from infected black pepper through somatic embryogenesis and meristem-tip culture , Crop Protection , 103., 1., Elsevier.