ICAR - National Bureau of Plant Genetic Resources (NBPGR), New Delhi http://nahep-caast.iari.res.in/. 18 From the field to the herbarium of cultivated plants "Hidden garden of plant genetic resources".
Characterization of Plant Genetic Resource for Genomics-assisted Crop Improvement
Role of ICAR-NBPGR in PGR Management
Systematic explorations were carried out in the most remote parts of the country, including Mon district of Nagaland;. A large number of genomic resources in the form of genomic and genic SSRs have been generated.
Some Important Plant Resources from Western Ghats
R. Yadav
Some of the rare endemic plants that have found a place in the garden are Ceropegia juncea and Frerea indica, the latter of which is critically endangered. It is necessary to know the autecology of at least critically endangered plant species in the country.
Exploration and Germplasm Collection: Status, Priorities and Future Thrust
This activity was more systematized after the establishment of the National Bureau of Plant Genetic Resources in 1976. This activity was more systematized after the establishment of the National Bureau of Plant Genetic Resources (NBPGR), New Delhi in 1976.
Germplasm Collecting: Logistics and Tactics
For this purpose, the detailed route maps (state and district) should be obtained for reference. The observations of variability parameters should be recorded along with photographs for documentation and publication.
International and National Regulation Governing the Use of Plant Genetic Resources
Applications for new varieties must be made with full passport details of all genetic resources used in the development of new material. Dua RP, P Brahmi and BS Dhillon (2004) International Treaty on Plant Genetic Resources for Food and Agriculture: A Review.
Introduction and Exchange of Plant Genetic Resources
Pursuant to the provisions of the Convention on Biological Diversity (CBD), the Government of India enacted legislation called the Biological Diversity Act (BDA), 2002 and also notified the Biological Diversity Rules, 2004, which suggest that no person from outside India or a body corporate, association, organization incorporated or registered in India with non-Indian participation in its share capital or management may access any biological resources or knowledge associated with research, commercial exploitation, bio-prospecting or bio-exploitation without prior approval of National Biodiversity Authority (NBA). For Annex 1 crop requests under ITPGRFA and FAO designated accessions of CG Centers located in India (ICRISAT), ICAR-NBPGR facilitates the process for approval by the Competent Authority viz.
Principles of Plant Quarantine and National Quarantine Set-up in India
C. Dubey* and Kavita Gupta
- Minimal impact: Phytosanitary measures shall be consistent with the pest risk involved, and shall represent the least restrictive measures available which result in the minimum
- Equivalence: Countries shall recognize as being equivalent those phytosanitary measures that are not identical but which have the same effect
- Technical authority: Countries shall provide an official Plant Protection Organization
- Managed risk: Because some risk of the introduction of a quarantine pest always exists, countries shall agree to a policy of risk management when formulating phytosanitary
- Emergency action: Countries may, in the face of a new and/or unexpected phytosanitary situation, take immediate emergency measures on the basis of a preliminary pest risk
Harmonization: Phytosanitary measures are based, whenever possible, on international standards, guidelines and recommendations developed under the IPPC. The Directorate of Plant Protection, Quarantine and Storage (DPPQS) of the Ministry of Agriculture and Farmers Welfare is the apex authority for the implementation of Plant Quarantine Regulations and the Plant Quarantine (Importation into India Regulation) Order, 2003 is the basis for the operation of the Directorate. This department functions under the delegated powers of the DPPQS (National Plant Protection Organization of India) Plant Quarantine Order, 2003.
Quarantine Procedures for Exchange of Plant Genetic Resources
The seeds are boiled in lacto-phenol solution (phenol, lactic acid, distilled water and glycerin in the ratio of 2:2:2:1 respectively) for 1-2 hours, depending on the hardness of the seeds. In case post-entry quarantine examination of the imported material is required, it is done at Quarantine Greenhouse Facility, at ICAR-NBPGR, New Delhi and its Regional Station, Hyderabad. These interceptions, especially of pests and their variability not yet reported from India [Category (i) and (ii)] indicate the importance of quarantine to prevent the introduction of destructive exotic pests. The categories (iii) and (iv) pests were not expected in the sample according to the literature-based PRA and since no records of the pest/host are available, their presence is unexpected and therefore important from a quarantine point of view.
Plant Genetic Resources - Conservation and Management
The principles governing conservation procedures are to preserve the viability, vitality and genetic integrity of the germplasm. Drying of the received germplasm under controlled conditions (drying rooms/cupboards are maintained at 15°C and 15% RH). Monitoring of preserved germplasm (10 years for most crops, except for oilseeds, where monitoring is carried out after 5-6 years).
Plant Tissue Culture Interventions for Management of Plant Genetic Resources
The flow of germplasm for in vitro conservation (intermediate term) in the in vitro Active Genebank (IVAG) and in vitro cryopreservation (long term) in the in vitro Base Genebank (IVBG) is graphically depicted in Figure 2. Agrawal A, S Singh, EV Malhotra, DPS Meena and RK Tyagi (2019) In vitro preservation and cryopreservation of clonally propagated garden species. Pandey R, N Sharma, A Agrawal, S Gupta, A Jain and RK Tyagi (2015) In vitro preservation and cryopreservation of vegetatively propagated crop germplasm.
Principles and Applications of Cryopreservation
The morphology of the embryo axes, shoot tips, meristematic tissues and dormant buds should be studied in advance for rapid and complete explant excision for various experimentation. The weight of the sample container should be comparable to the explant to be weighed. Regular monitoring of the sample should be done to ensure high viability and low moisture content.
Characterization and Evaluation of Plant Genetic Resources
Gangopadhyay 1 *, Kuldeep Tripathi and Pooja Kumari 2
It generally starts with a large number of germplasm from the National Genebank or germplasm already identified by characterization and preliminary evaluation or based on passport data, and is carried out in a single location adjusted for agronomic traits. The evaluation is carried out with an appropriate experimental design, depending on the objective and the size of the germplasm, with at least three examinations, viz. Generally, characterization and preliminary assessment is done by germplasm curator/scientists; further evaluation or detailed evaluation is mostly done by breeders to obtain additional information.
Evaluatio n
This is one of the effective methods to explore PGR for climate change adaptive properties specific germplasm. Brown (1989) suggested that it should be no more than 10 percent of the entire collection and always less than 2000 records. Most of the core collections developed were 5–20% the size of the collection from which they were established.
Pre Breeding for Effective Use of Plant Genetic Resources
New cultivars incorporating genes from wild relatives were also reported to be in the pipeline. Use of cultivated wild relatives in the last 20 years in released cultivars of 13 crops of international importance (Reem and Toby, 2017). Hajjar Reem and Toby Hodgkin (2017) The use of wild relatives in crop improvement: A survey of developments over the past 20 years.
Advances in Plant Genetic Resources Evaluation
So, the examination of germplasm for biotic stresses should be accompanied by the identification of the race / strain / isolate / biotype of the pest. Advanced field testing of selected toleranced parts in the field should be done for at least two years. For recording data on viral diseases, incidence (number of plants showing symptoms over total plant number) and average disease severity (number of leaves showing symptoms or area covered over total number of leaves on a single plant and averaged from at least five plants) should be considered.
Role of Genomics in PGR Management
Page | 121 Next-generation sequencing technologies have revolutionized research by allowing an economical elucidation of the genome of any organism. At NBPGR, the generation, validation and use of genomic resources is the primary objective of the Division of Genomic Resources. Further characterization of the minicore in a nuclear collection applied using the focused identification of germplasm strategy (FIGS) has been accelerated using next-generation sequencing technologies.
DNA Fingerprinting of Crop Varieties
Page | 126 an authentic, representative sample of the variety and, where possible, must be obtained from the sample of the variety used for examination for purposes of Plant Breeders. The plant material from which the samples are taken must be traceable if some of the samples later turn out not to be representative of the variety. With regard to the representation of the variety, consideration must be given to the method of propagation ie.
Plant Genetic Resources Informatics
Genebank curators, breeders and students, as well as policy makers and funding agencies can rely on PGR Informatics tools and applications to plan and implement activities related to PGR management and application. The following sections list major PGR informatics applications developed at NBPGR and used by researchers worldwide. PGR Informatics' products and services at NBPGR are a culmination of the efforts and support of several individuals in the Bureau over a long period of time.
From Field to the Herbarium of Cultivated Plants “The Hidden Garden of Plant Genetic Resources”
The herbarium specimens in general and especially of the wild species and wild relatives should be collected to aid in identification. The hot air flowing through the corrugated plates (placed between the blotters) facilitates the drying of the samples. NHCP has begun georeferencing herbarium specimens of the selected taxa and plans to continue.
Detection and Identification of Pests in Exotic Germplasm including Transgenics
Detection and Identification of Plant Viruses
Real-time RT-PCR: Real-time PCR monitors the fluorescence emitted during the reaction as an indicator of amplicon production during each PCR cycle (ie, in real time) as opposed to the endpoint detection. The advantages of Real-time PCR/ Real-time RT-PCR are that it is not affected by non-specific amplification; gain can be monitored in real time; no post-PCR processing of products (high throughput, low contamination risk); ultra-fast cycling (30 minutes to 2 hours); requires 1000 times less RNA. The intercept includes 19 destructive viruses not yet reported from India namely Barley stripe mosaic virus, mild bean mosaic virus, bean mottle virus, broad bean melt virus, broad bean spot virus, broad bean sand mosaic virus, Cherry leaf roll virus, Cowpea mottle virus, Cowpea severe mosaic virus, Dioscorea latent virus, Garlic virus C, High plains virus, Maize chlorotic mottle virus, Pea enation mosaic virus, Peanut sunt virus, Pepino mosaic virus, Raspberry ringspot virus, Tomato ringspot virus and Wheat streak mosaic virus.
Detection and Identification of Fungi and Bacteria
Method of soaking seeds: For detecting bunt (Tilletia barclayana) in rice, the seeds are soaked overnight in 0.2 percent sodium hydroxide and examined. For the detection of bacteria, the surface-sterilized seeds are placed on nutrient agar medium in Petri plates and incubated at 22–25 °C. Several selective/semi-selective media have been developed and used for the detection of specific bacteria associated with seeds.
SECTION III: Detection and Identification of Plant Parasitic Nematodes
Passive dispersal is a long-distance movement of nematodes by various agents such as wind, water, field equipment, planting material, etc. In addition to these natural means of dispersal, anthropogenic factors are very common for long-distance dispersal of nematodes. It includes careful observation of microplasma, extraction of nematode stages, identification and proper control of them.
Detection and Identification of Insects and Mites
Placement of mites - Slide holders are prepared for the intercepted mites, which are then identified with identification keys. Some mites are semi-transparent or transparent, which are painted before installation for easier identification. Based on morphological and key features, pests are identified with an identification key.
Detection and Identification of Weeds
Page | 167 Bhalla S, VC Chalam, B Singh, K Gupta and SC Dubey (2018) Biological conservation of plant genetic resources in India: Role of plant quarantine. Chalam VC and RK Khetarpal (2008) A critical appraisal of the challenges in excluding plant viruses during transboundary movement of seeds. ICAR-National Bureau of Plant Genetic Resources and Ministry of Environment, Forests and Climate Change, New Delhi, India.
Conservation Procedures for Seed Genebank
Flow Chart for Seed Germplasm Conservation
Drying of germplasm
For long-term storage, seeds of each accession should be dried to a moisture content of 3-7%, depending on the species. If the moisture content of the received sample appears to be below 17%, the connections may be subjected directly to oven drying. If the seeds required pre-drying, the moisture content is calculated twice, i.e. once after pre-drying and secondly using the oven method.
Testing for Seed Viability
Seeds are sown on paper and petri dishes are incubated in incubators at high humidity and appropriate/optimal temperature as recommended by ISTA (Table 1). Seedlings are scored on the day fixed for the final count and are classified as normal seedlings, abnormal seedlings, fresh ungerminated seeds and dead seeds. ISTA has recommended specific dormancy breaking treatments for each species based on the nature of dormancy.
Data sheet for recording Seed Vigour results
Data sheet for recording Electrical Conductivity
Quick Viability Testing
Testing can be done at room temperature, but it will take longer to stain. Temperatures above 40°C should not be used. Among colored seeds, seeds with bright red coloration are fully viable, while partially colored seeds can produce either normal or abnormal seedlings. Seeds completely and uniformly stained (2) seeds showing minor unstained areas on cotyledon opposite deeply stained cotyledon tip and parts of cotyledon (3) Seeds showing necrotic tissue towards cotyledon tip and in center of cotyledon (4) seeds completely unstained .
