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Improvement of Medicinal Plants: Challenges and Innovative Approaches

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Senthil Natesan
Senthil Natesan

This document discusses breeding efforts to develop improved varieties of the medicinal plant Ashwagandha (Withania somnifera). It describes the development of two promising lines, DWS 132 and DWS 135, as well as a dwarf pure line called DWS 327. A new procumbent plant type was developed called DWS-6. For the first time, a male sterile line was developed in ashwagandha. Growth studies showed active ingredients decrease with maturity. Some identified descriptors for distinctness, uniformity and stability include plant height, type, branching pattern and root branching.

Science
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P. Manivel Directorate of Medicinal and Aromatic Plants Research, Boriavi-387310, Anand, Gujarat *Email: manivelp@yahoo.com; 9429543209 1
India’s strength in Herbal TechnologyTHEINDIANFLORA(ca17500species) INDIANSYSTEMSOFMEDICINE
Climbers 12% Others 3% Shrubs 20% Herbs 32% Trees 33%Wild 90% Cultivation 10% Medicinal Plants by Source in India Distribution of Medicinal Plants by Habits in India
Roots 30% Bark 14%Whole plant 16% Flowers 5% Leaves 6% Fruits 10 % Rhizome 4% Stems 6% Seeds 7% Wood 3%
5
0 1000 2000 3000 4000 5000 2002 2020 2030 2050 60 1200 1740 5000 US$Billions Year  80% of the world population depend on traditional systems of medicines for primary health care  China and India are the worlds leading exporters of MAP based drugs.
 Factors  Population  Food habit  Climate change  Life style  Genetic  Pollution • Major health problems Cancer, Hypertension, Diabetes, Malarial, Viral Dengue, Chikengunea and others Modern medicineTraditional systems of Medicine
M&AP Distribution over 10 Biogeographic Regions of India
AREA OF ACTIVITIES • Crop Improvement • Crop Production • Crop Protection • Quality Assessment • Post Harvest Management CROPS Working on 35 species
Aconitum heterophyllum Adenathera pavonia Amorphophalus campanulatus (Elephant foot yam) Barleria prionitis, B. Cristata Baliospermum montanum Berberis aristata Chonemophora fragrance (murva) Cinnamomum sulphuratum Commiphora wightii Coscinium fenestratum Dactylorhiza hataqgeria Decalepis hamiltonii Dioscorea deltoidea Ephedra gerardiana Gentiniana kurroo Leptadenia reticulata Lilium polyphyllum Malaxis muscifera Myristica malabarica Oryxylum indicum Picrorhiza kurrooa (Kutki) Podophylum hexandrum Polygonatum verticillatum Pterocarpus santalinus (Red sanders) Raulvolfia serpentina (Sarpagandha) Rheum australe Saraca asoca Swertia chirayita Taxus wallichiana Tinospora malabarica Tinospora cordifolia Trichospus zeylanicus Valeriana hardwickii Vateria indica Withania coagulans
Sr. no Common name Botanical name 1. Amla Emblica officinalisGaertn 2. Ashok Saraca asoca (Roxb.) de wilde 3. Ashwagandha Withania somnifera (Linn.) Dunal 4. Atees Aconitum heterophyllum Wall. ex Royle 5. Bel Aegle marmelos (Linn) Corr. 6. Bhumi amlaki Phyllanthus amarus schum &Thonn. (P. niruri Linn.) 7. Brahmi Bacopa monnieri (L.) Pennell 8. Chandan Santalum album Linn. 9. Chirata Swertia chirata Buch-Ham. 10. Daruhaldi Berberis aristata DC. 11. Gudmar Gymnema sylvestre R. Br. 12. Guduchi Tinospora cordifolia Miers. 13. Guggal Commiphora wightii (Arn.) Bhandari 14. Isabgol Plantago ovata Forsk. 15. Jatamansi NardostachysJatamansi DC. 16. Kalihari Gloriosa superba Linn. 17. Kalmegh Andrographis paniculataWall. Ex Nees Sr. no Common name Botanical name 18. Kesar Crocus sativus Linn. 19. Kokum Garcinia indicaChois. 20. Kuth Saussurea costusC. B. Clarke (S.lappa) 21. Kutki Picrorhiza kurroa Benth ex Royle 22. Makoy Solanum nigrum Linn. 23. Mulethi Glycyrrhiza glabra linn. 24. Pathar chur (Coleus) Coleus barbatus Benth. 25. Pippali Piper longum Linn. 26. Safed Musli Chlorophytum arundinaceum Baker (C. borivillianum) 27. Sarpgandha Rauvolfia serpentina (L.) BENTH. EX KURZ 28. Senna Cassia angustifoliaVahl. 29. Shatavari Asparagus racemosusWilld. 30. Tulsi Ocimum sanctum Linn. 31. VaiVidang Embelia ribes Burm. f. 32. Vatsnabh Aconitum ferox wall. 13
 Plantago ovata  Cassia aungustifolia  Withania somnifera  Aloe barbadensis  Asparagus recemosus  Papaver somniferum  Chlorophytum borivilianum  Piper longum  Plumbago rosea  Solanum nigrum  Centella asiatica  Gymnema sylvestre  Aconitum heterophyllum  Ocimum sanctum  Saraca asoca  Bacopa monnieri  Swertia chirayita  Valeriana jatamansi  Hypericum perforatum  Leptadenia reticulata  Convolvulus microphyllus  Andrographis paniculata  Sida cordifolia  Acorus calamus  Rawvolfia serpentine  Tinospora cordifolia  Podophylum hexandrum  Aloe barbadensi
PRESENT BREEDING STATUS MEDICINAL PLANT VARIETIES
AICRP on MAPB & DMAPR (ICAR) CIMAP (CSIR) No. of species Total number of varieties Medicinal Plants 22 33 Aromatic Plants 6 7 No. of species Total number of varieties Medicinal Plants 20 25 Aromatic Plants 18 42
Sl No Crop Variety Developed by Year of release Medicinal Plants 1 Chlorophytum borivilianum (Safed musli) Jawahar Safed musli 405 AINRP on M &AP, Jawaharlal Nehru Krish VishwaVidyalaya, Mandsaur 2004 2 Catharanthus roseus (Periwinkle) Prabhat AINRP on M&AP, CCS Haryana Agricultural University, Hisar 2002 3 Cassia angustifolia Vahl. (Senna) ALFT-2 AINRP on M &AP, Gujarat Agricultural University, Anand 1989 4 Diascoria floribunda Mart. & Gal. (Yam) FB(C)-1 Indian Institute of Horticulture Bangalore 1974 5 Arka Upakar Indian Institute of Horticulture, Bangalore 1980 6 Digitalis lanata Ehrh. (Foxglove) D 76 AINRP on M&AP,Y.S.Parmar University of Horticulture and Forestry, Nauni Solan 1991 7 Glaucium flavum (Yellow Horned Poppy) H 47-3 AINRP on M&AP,Y.S.Parmar University of Horticulture and Forestry, Nauni Solan 1991 8 Glycyrrhiza glabra L. (Liquorice) Haryana Mulhatti-1 AINRP On M&AP, CCS Haryana Agricultural University, Hisar 1989 9 Hyoscyamus muticus L. (Egyptian Henbane) HMI-80-1 AINRP on M&AP, College of Agriculture, Indore _
Sl No Crop Variety Developed by Year of release Medicinal Plants 10 Lepidium sativum L. (Cress) GA 1 AINRP M &AP, Center, Gujarat Agricultural University, Anand 1998 11 Rauvolfia serpentine Benth. Ex Kurz. (Sarpagandha) RI-1 AINRP on M&AP, College of Agriculture, Indore _ 12 Papaver somniferum L. (Opium Poppy) Jawahar Aphim 16 AINRP on M&AP, College of Agriculture, Mandsaur 1984 13 Kirtiman AINRP on M&AP,ND University of Agriculture andTechnology, Faizabad 1990 14 Jawahar Opium 539 AINRP on M&AP, College of Agriculture, Mandsaur 1997 15 Jawahar Opium 540 AINRP on M&AP, College of Agriculture, Mandsaur 1998 16 Chetak Aphim AINRP on M&AP, Rajasthan Agricultural University, Udaipur 1994 17 Trishna NBPGR, New Delhi NOP-1 AINRP on M&AP, ND University of Agriculture andTechnology, Faizabad
PRESENT BREEDING STATUS GERMPLASM
Conservation Method Holding Important MAP with sizeable variability LTS (cultivated) 2320 Papaver, Plantago, Andrographis, Asparagus, Chlorophytum, Mucuna, Psoralea, Withania, Ocimum etc. LTS (wild) 2316 (652 sp.) Terminalia, Tribulus, Rheum, Helecteris, Brynopsis, Mimosa, Morinda, Cassia, Zanthoxylum, Holarrhena, Boerhaavia, etc. Status of MAP in National Genebank
Sl. No. Species Total 1. Aloe spp. 55 2. Andrographis paniculata 60 3. Asparagus spp. 88 4. Cassia angustifolia 222 5. Chlorophytum borivilianum 54 6. Commiphora spp 225 7. Cymbopogon martinii 07 8. Desmodium gangeticum 52 9. Gymnema sylvestre 43 10. Plantago spp. 91 11. Tinospora cordifolia 52 12. Withania somnifera 142 TOATAL 1091
22 INGR No.04113: Fleshy roots are long (>10 cm) with blunt end and dark colour, arranged compactly in the bunch (converged type) of Safed musli (Chlorophytum borivilianum) INGR No.04114: Fleshy roots are short (<10 cm) with blunt end and light colour, arranged in wide angles from the axis (diverged type) and with excellent storage quality of Safed musli (Chlorophytum borivilianum) INGR No.06023 : Superior gel rich clone (2191.18 g/ plant) of Aloe INGR No.06024 : Superior aloin-A rich clone (26.13%) of Aloe INGR No.06025 : High starch containing clone (13.32%) of Giloe INGR No.07041 : Compact plant type with high andrographolide content of Kalmegh DWS 327 (INGR 11026): A dwarf (<30 cm) Ashwagandha (Withania somnifera) pure line DWS-6 (INGR 13047) : A new Procumbent type plant of Ashwagandha (Withania somnifera) DGS 22 (INGR 13041): a high fruit producing Madhunashini germplasm. DPO 14 (INGR 11035): An early maturing (85-90 days) and high harvest index Isabgol (Plantago ovata) mutant DWS 296-4 (INGR14010): A golden yellow colour leaf mutant of Isabgol (Plantago ovata Forsk) Elite PGRs registered with NBPGR
Conservation In situ Community Conservation Ex situ  National Parks  Protected Areas  Biosphere Reserves  World Heritage Sites (Forest Departments)  Medicinal plants conservation Areas  Medicinal Plants Development Areas In situ Ex situ On- farm On-farm •Seed gene banks •Botanical or Herbal gardens •Home Herbal Gardens •Community herbal gardens •Medicinal plants conservation parks •Cultivation of MAPs •Field gene banks •In vitro repository - Land races - Sacred groves - Folk Varieties (Tribal and Rural families) 23
BREEDING OF Withania somnifera
Antioxidant Adaptogen Aphrodisiac Liver tonic Anti-inflammatory agen Astringent Treat ulcers Bacterial infection Venom toxins Cancer Astringent is a substance which causes biological tissue to contract or draw together Adaptogens help balance our bodily functions via the neurotransmitters of the brain. (sexual stimulant)
DWS 135DWS 132 DWS 132 and DWS 135: Promising ashwagandha lines entered in to IVT-I (2013-14)
DWS 327: A dwarf (<30 cm) ashwagandha (Withania somnifera) pure line (INGR 11026) DWS 327 JA 134
DWS 327 DWS-6: A new Procumbent type plant of ashwagandha (Withania somnifera) (INGR 13047)
First time in ashwagandha male sterile line has been developed
Pollen germination in MS stigma Pollen tube growing Pollen tube reaching to ovary No Pollen germination on selfing MS Pollen MS anther
Pollinators in male sterile plants Ants House flies Honey Bees 3rd day after pollinationMeiosis was normal in MS
Growth study in Ashwagandha Very slow in the initial period up to 75 days Steadily increased from 90 to 150 DAS. The initial dry matter increase – due root and leaves  After 90 days it was due to fruits and leaves.
 Young roots had more 12-deoxywithanostramonolide and Withanolide A content per unit root weight  During maturity the content of active ingredients decreased. Chemical changes during growth 12-deoxywithanostramonolide (%) Withanolide A content (%) 0 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0 1 2 3 4 5 6 7 JA-20 JA 134 Linear (JA 134) Linear (JA-20) 90 105 120 135 150 160 0 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0.08 0 1 2 3 4 5 6 7 JA-20 JA 134 Linear (JA 134) Linear (JA-20) 90 105 120 135 150 160
Some DUS descriptors identified Dwarf Extra dwarf Trait Descriptor • Plant height • Tall (> 90 cm) • Medium (30- 90 cm) • Dwarf (< 30 cm) • Plant type 1. Erect 2. Semi erect 3. Spreading 4. Prostrate
Extra dwarf Trait Descriptor • Branching pattern • No branches • Few (2-3) branches • More branches (4-6) in the bottom • Profuse branches (branch in each node) • Root branching pattern • No branches • Few (1-2) branches • More branches (3-4) • Profuse branches (>5)
Variability for plant type Broad leaf Narrow leaf Open type More branching Compact plantErectWhitish leaf
Extra dwarf Trait Descriptor • Calyx attachment with berry • Loosely attached • Moderately attached • Tightly attached • Calyx tip • Fully open • Semi open • Closed • Berry (Calyx) shape • Oblong • Roundish • Calyx covering berry • Open • Closed
Trait Descriptor • Leaf shape • Ovate • Triangle • Elongate • Leaf margin • Straight • Wavy •Leaf colour • Light green • Dark green •Leaf size • Broad • Narrow •Leaf Curling • Downward curling • Upward curling
Variability for leaf characters Moderate wavy More wavy Curved leaf Straight margin Long leaf
Some morphologically distinct types Extra dwarf Open type calyx Cluster flowers Less fibrous rootProcumbent type Less flowers More hairy Leaf Colour
Variability for berry color Yellow Red Orange Creamy white
DWS 326DWS 6
Jain et al., 2009: J Plant biochemistry and Biotechnology,18(2):249-252 Micro propagation of Withania coagulans Dunal:A critically endangered medicinal plant Induction of shoot bud from nodel explants Proliferation of shoots Root initiation Similar banding pattern of tissue cultured plants
MDendrogram constructed using 27 RAPD markers AP 4 Lane: 1 100bp ladder and 2-25 withania accessions
45
84 Isabgol germplasm accessions are being maintained. Collection, purification & maintenance of germplasm-ISABGOL Six Plantago species P. indica, P. arinaria, P. psyllium, P. lanceolata, P. serrana, and P. carnopus
Standardize the hybridization procedure No. of floweers emasculated Seed obtain ed % crossing success E & S 203 12 5.91 E & P 212 73 34.43
Plantago ovata GERMPLASM
DPO 4 GI 2 DPO 4 DPO 1 DPO 1 TWOVarieties Developed DPO 4 had 30, 15 and 14% higher yield than GI 2, Niharika and local check, respectively DPO 1 had 21, 19 and 14% higher yield than GI 2, Niharika, GI 2, and local check, respectively
Early maturing mutant (75-80 d) M2 Flower initiation in 30 days Matures in 80-85 days 100% Flowering in 35-40 days DPO 14:An early maturing (85-90 days) and high harvest index isabgol (Plantago ovata) mutant (INGR 11035)
Golden yellow leaf mutant (INGR 14010) DPO 296-4: A golden yellow leaf colour mutant
Closed flower mutant Whitish leaf base and stem Short leaf with twisted leaf tip (Dark green) Short leaf with light green colour
Short (> 10 cm)leaf with straight leaves Yellowish green leaves Narrow leaf
Short (> 8 cm) stem -prostrate Inflorescence within the canopy
Narrow leaf ControlDiseased ControlDiseased Downey mildew resistance sources identified
ControlDiseased
DPO 185 (resistant) DPO 144, DPO 145, DPO 185 and DPO 333-2 Downey mildew resistance sources identified
Physiological study of Isabgol mutants/lines Morning Afternoon Morning Afternoon -25 -20 -15 -10 -5 0 Morning Afternoon Morning Afternoon DPO 18 DPO 22 DPO 30 DPO 38 DPO 44 DPO 61 DPO 69 DPO 70 DPO 72 DPO 78 DPO 113 DPO 126 DTPO 15 DPO 136 DTPO 21 DPO 144 DPO 162 DPO 181 DPO 14 DPO 185 DPO 189 DPO 195 DPO 200 DPO 204 DPO 206 DPO 216 DPO 230 DPO 231 DPO 232 DPO 233 DPO 240 DPO 241 DPO 256 DPO 264 DPO 266 DPO 267 DPO 269 DPO 270 DPO 274 DPO 280 DPO 281 DPO 282 DPO 283 DPO 296 DPO 307 DPO 311 DPO 312 DPO 332 DPO 334 DPO 352 DPO 356 DPO 362 DPO 371 DPO 395 DPO 396 DPO 415 DPO 422 DPO 431 DPO 433 DPO 437 Leaf water potential of Isabgol lines at saturated soil moisture and water deficit conditions. Irrigated Drought
Tetraploids were developed in isabgol Out of 17 lines were screened for its ploidy level (diploid vs tetraploid) using FACS analysis and tetraploid has been confirmed in 97 lines File:DTPO4-2op Date:10-03-2011 Time:12:15:18 Particles:1068 Acq.-Time:236 s 0 200 400 600 800 1000 0 20 40 60 80 100 FL4 DAPI counts 0 200 400 600 800 1000 0 20 40 60 80 100 FL4 DAPI counts partec PAS File:Control 1 (DPO 1) Date:10-03-2011 Time:11:26:27 Particles:770 Acq.-Time:349 s 0 200 400 600 800 1000 0 20 40 60 80 100 FL4 DAPI counts 0 200 400 600 800 1000 0 20 40 60 80 100 FL4 DAPI counts partec PAS FACS analysis 2x 4 x 2x File:DTPO2-1 Date:09-02-2011 Time:17:03:48 Particles:1027 Acq.-Time:255 s 0 200 400 600 800 1000 0 20 40 60 80 100 FL4 DAPI counts 0 200 400 600 800 1000 0 20 40 60 80 100 FL4 DAPI counts partec PAS 4x2x Diploid Tetraploid TetraploidDiploid
Raw reads from NCBI Quality filteration / trimming of raw reads Prineq-lite Clean high quality reads Denovo transcriptomeAssembly Trinity Assembler AssembledTranscripts Evaluation of Assembly CDS prediction Annotation SSR prediaction GO analysis Blast2GOBlastX MISA Primer3 Primer designing CLC Genomic Workbench InhouseTranscript Workflow ofTranscriptome analysis
 The raw data Plantago sample was downloaded from NCBI (http://www.ncbi.nlm.nih.gov/sra/?term=Plantago).  Raw data was filtered using Prineq-lite.  Adapter contamination and reads with low quality value i.e. an average QV less than 20 (QV < 20) were filtered.  The high quality data of Plantago sample was assembled with CLC genomics workbench on default parameters (mismatch cost = 2, Insertion/Deletion cost = 3) for denovo assembly.  A total of 23,586 transcript contigs were obtained for Plantago sample.
 The functional annotation was performed using 23,586 transcript contigs of Plantago sample by aligning against NR database of NCBI using BLASTX.  The annotated transcript contigs were mapped on GO database which showed significant similarity with Solanum tuberosum followed by Vitis vinifera, Solanum lycopersicum and so forth.  The distribution of annotated transcript contigs in specific ontology domains indicate 4192 GO terms were assigned to Biological Processes, 2352 GO terms to Molecular Functions and 5309 terms to Cellular Components.
1=100 bp ladder 2=DPO-185 3=DPO-14 4=Club mutant 5=EC 124345 6=EC 427062 7=G-I-3 8=H -I-29 9=DM -4 10=MIB-121 11=DM -1 12=P -79 13=PB-6-3 14=AMB -2 15=DM -2 16=DM -11 17=P -1 18=MALE STERILE 19=DM-6 20= DM-5 31= MIB-124 32=DM-7 33= P-33 34=JI-9 35= PB-10 -4 36= MIB-2 37=UR-188 38=MIB-4 39=MIB-122 40= PS-19 51=A-24 52 H-I-9 53= DM-3 54= RI-88 55= Mutant 56= H-I-6 57= RI-9809 58= RI-157 59= HI-34 60= PB-96 61=100 bp ladder 62= P-44 63= MIB-2 64= PB-6-1 65= RI-49 66= Sel -10 67= MI-4 68=RI-158 69= P-6 70= RI-130 71= DM-10 72= DM-8 73= G-I-1a 74=G-I-2 75=P.Indica 76=P.arineria 77=P.psyllium 78=P.Lanceolata 79= P.Cerina 80=P.Carnopus 21=100 bp ladder 22=AM -5 23=Palampur-3 24= MSB-8 25= J-1B 26=RI-129 27=PS-17 28= RI-89 29= PB-3-1 30=PB-31 41=100 bp ladder 42= MIB-123 43= PB-80 44= Palampur -2 45= RI-149 46= PB-62 47= MIB-125 48= RI-9808 49= RI-153 50= RI-156 Xdapoem21 marker Polymorphism study in different germplasm of Isabgol
Asparagus recemosus germplasm
Gymnema sylvestre germplasm
DGS 22 DGS 22: a high fruit producing Madhunashini germplasm (INGR 13041)
Desmodium gangeticum
DAA 1 DAA 2 DAA 3 DAA 4 DAA 5 Asparagus adscendens
DAR 1 DAR 2 DAR 3 DAR 4 DAR 5 Asparagus racemosus
Desmodium gangeticum DDG 1 DDG 2 DDG 3 DDG 4
Gymnema sylvestre (22 Acc.) DGS 1 DGS 2 DGS 3 DGS 4 DGS 5 DGS 6
• Extent of genetic variability • Knowledge of reproductive biology • Information on the genetics of agronomic traits of interest • Information on the genetics of pathways of secondary metabolism. • Breeding strategies • Availability of efficient analytical techniques for quality analysis
Disease Species to be targeted Hypertension Guggul (Commiphora weightiii) Sarpagandha (Rauvolfia serpentina) Terminalia species Diabetes Mamejo (Enicostema axillare) Gurmar (Gymnema sylvestere) Cancer Kokum (Garcinia) species Annona Dengue Papaya (Carica papaya) Immunomodulation Ashwagandha (Withania somnifera) Kalmegh (Andrographis paniculata Malaria Artimesia (Artemisia annua) Women health Shatavari (Asparagus racemosus) Asoca (Saraca asoca) Parkinson’s or Alzheimer disease Velvet beans (Mucuna pruriens) Laxative Isabgol (Plantago ovata) Senna (Cassia angustifolia)
THANKS

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Improvement of Medicinal Plants: Challenges and Innovative Approaches

  • 1. P. Manivel Directorate of Medicinal and Aromatic Plants Research, Boriavi-387310, Anand, Gujarat *Email: manivelp@yahoo.com; 9429543209 1
  • 2. India’s strength in Herbal TechnologyTHEINDIANFLORA(ca17500species) INDIANSYSTEMSOFMEDICINE
  • 5. 5
  • 6. 0 1000 2000 3000 4000 5000 2002 2020 2030 2050 60 1200 1740 5000 US$Billions Year  80% of the world population depend on traditional systems of medicines for primary health care  China and India are the worlds leading exporters of MAP based drugs.
  • 7.  Factors  Population  Food habit  Climate change  Life style  Genetic  Pollution • Major health problems Cancer, Hypertension, Diabetes, Malarial, Viral Dengue, Chikengunea and others Modern medicineTraditional systems of Medicine
  • 8.
  • 9.
  • 10. M&AP Distribution over 10 Biogeographic Regions of India
  • 11. AREA OF ACTIVITIES • Crop Improvement • Crop Production • Crop Protection • Quality Assessment • Post Harvest Management CROPS Working on 35 species
  • 12. Aconitum heterophyllum Adenathera pavonia Amorphophalus campanulatus (Elephant foot yam) Barleria prionitis, B. Cristata Baliospermum montanum Berberis aristata Chonemophora fragrance (murva) Cinnamomum sulphuratum Commiphora wightii Coscinium fenestratum Dactylorhiza hataqgeria Decalepis hamiltonii Dioscorea deltoidea Ephedra gerardiana Gentiniana kurroo Leptadenia reticulata Lilium polyphyllum Malaxis muscifera Myristica malabarica Oryxylum indicum Picrorhiza kurrooa (Kutki) Podophylum hexandrum Polygonatum verticillatum Pterocarpus santalinus (Red sanders) Raulvolfia serpentina (Sarpagandha) Rheum australe Saraca asoca Swertia chirayita Taxus wallichiana Tinospora malabarica Tinospora cordifolia Trichospus zeylanicus Valeriana hardwickii Vateria indica Withania coagulans
  • 13. Sr. no Common name Botanical name 1. Amla Emblica officinalisGaertn 2. Ashok Saraca asoca (Roxb.) de wilde 3. Ashwagandha Withania somnifera (Linn.) Dunal 4. Atees Aconitum heterophyllum Wall. ex Royle 5. Bel Aegle marmelos (Linn) Corr. 6. Bhumi amlaki Phyllanthus amarus schum &Thonn. (P. niruri Linn.) 7. Brahmi Bacopa monnieri (L.) Pennell 8. Chandan Santalum album Linn. 9. Chirata Swertia chirata Buch-Ham. 10. Daruhaldi Berberis aristata DC. 11. Gudmar Gymnema sylvestre R. Br. 12. Guduchi Tinospora cordifolia Miers. 13. Guggal Commiphora wightii (Arn.) Bhandari 14. Isabgol Plantago ovata Forsk. 15. Jatamansi NardostachysJatamansi DC. 16. Kalihari Gloriosa superba Linn. 17. Kalmegh Andrographis paniculataWall. Ex Nees Sr. no Common name Botanical name 18. Kesar Crocus sativus Linn. 19. Kokum Garcinia indicaChois. 20. Kuth Saussurea costusC. B. Clarke (S.lappa) 21. Kutki Picrorhiza kurroa Benth ex Royle 22. Makoy Solanum nigrum Linn. 23. Mulethi Glycyrrhiza glabra linn. 24. Pathar chur (Coleus) Coleus barbatus Benth. 25. Pippali Piper longum Linn. 26. Safed Musli Chlorophytum arundinaceum Baker (C. borivillianum) 27. Sarpgandha Rauvolfia serpentina (L.) BENTH. EX KURZ 28. Senna Cassia angustifoliaVahl. 29. Shatavari Asparagus racemosusWilld. 30. Tulsi Ocimum sanctum Linn. 31. VaiVidang Embelia ribes Burm. f. 32. Vatsnabh Aconitum ferox wall. 13
  • 14.  Plantago ovata  Cassia aungustifolia  Withania somnifera  Aloe barbadensis  Asparagus recemosus  Papaver somniferum  Chlorophytum borivilianum  Piper longum  Plumbago rosea  Solanum nigrum  Centella asiatica  Gymnema sylvestre  Aconitum heterophyllum  Ocimum sanctum  Saraca asoca  Bacopa monnieri  Swertia chirayita  Valeriana jatamansi  Hypericum perforatum  Leptadenia reticulata  Convolvulus microphyllus  Andrographis paniculata  Sida cordifolia  Acorus calamus  Rawvolfia serpentine  Tinospora cordifolia  Podophylum hexandrum  Aloe barbadensi
  • 16. AICRP on MAPB & DMAPR (ICAR) CIMAP (CSIR) No. of species Total number of varieties Medicinal Plants 22 33 Aromatic Plants 6 7 No. of species Total number of varieties Medicinal Plants 20 25 Aromatic Plants 18 42
  • 17. Sl No Crop Variety Developed by Year of release Medicinal Plants 1 Chlorophytum borivilianum (Safed musli) Jawahar Safed musli 405 AINRP on M &AP, Jawaharlal Nehru Krish VishwaVidyalaya, Mandsaur 2004 2 Catharanthus roseus (Periwinkle) Prabhat AINRP on M&AP, CCS Haryana Agricultural University, Hisar 2002 3 Cassia angustifolia Vahl. (Senna) ALFT-2 AINRP on M &AP, Gujarat Agricultural University, Anand 1989 4 Diascoria floribunda Mart. & Gal. (Yam) FB(C)-1 Indian Institute of Horticulture Bangalore 1974 5 Arka Upakar Indian Institute of Horticulture, Bangalore 1980 6 Digitalis lanata Ehrh. (Foxglove) D 76 AINRP on M&AP,Y.S.Parmar University of Horticulture and Forestry, Nauni Solan 1991 7 Glaucium flavum (Yellow Horned Poppy) H 47-3 AINRP on M&AP,Y.S.Parmar University of Horticulture and Forestry, Nauni Solan 1991 8 Glycyrrhiza glabra L. (Liquorice) Haryana Mulhatti-1 AINRP On M&AP, CCS Haryana Agricultural University, Hisar 1989 9 Hyoscyamus muticus L. (Egyptian Henbane) HMI-80-1 AINRP on M&AP, College of Agriculture, Indore _
  • 18. Sl No Crop Variety Developed by Year of release Medicinal Plants 10 Lepidium sativum L. (Cress) GA 1 AINRP M &AP, Center, Gujarat Agricultural University, Anand 1998 11 Rauvolfia serpentine Benth. Ex Kurz. (Sarpagandha) RI-1 AINRP on M&AP, College of Agriculture, Indore _ 12 Papaver somniferum L. (Opium Poppy) Jawahar Aphim 16 AINRP on M&AP, College of Agriculture, Mandsaur 1984 13 Kirtiman AINRP on M&AP,ND University of Agriculture andTechnology, Faizabad 1990 14 Jawahar Opium 539 AINRP on M&AP, College of Agriculture, Mandsaur 1997 15 Jawahar Opium 540 AINRP on M&AP, College of Agriculture, Mandsaur 1998 16 Chetak Aphim AINRP on M&AP, Rajasthan Agricultural University, Udaipur 1994 17 Trishna NBPGR, New Delhi NOP-1 AINRP on M&AP, ND University of Agriculture andTechnology, Faizabad
  • 20. Conservation Method Holding Important MAP with sizeable variability LTS (cultivated) 2320 Papaver, Plantago, Andrographis, Asparagus, Chlorophytum, Mucuna, Psoralea, Withania, Ocimum etc. LTS (wild) 2316 (652 sp.) Terminalia, Tribulus, Rheum, Helecteris, Brynopsis, Mimosa, Morinda, Cassia, Zanthoxylum, Holarrhena, Boerhaavia, etc. Status of MAP in National Genebank
  • 21. Sl. No. Species Total 1. Aloe spp. 55 2. Andrographis paniculata 60 3. Asparagus spp. 88 4. Cassia angustifolia 222 5. Chlorophytum borivilianum 54 6. Commiphora spp 225 7. Cymbopogon martinii 07 8. Desmodium gangeticum 52 9. Gymnema sylvestre 43 10. Plantago spp. 91 11. Tinospora cordifolia 52 12. Withania somnifera 142 TOATAL 1091
  • 22. 22 INGR No.04113: Fleshy roots are long (>10 cm) with blunt end and dark colour, arranged compactly in the bunch (converged type) of Safed musli (Chlorophytum borivilianum) INGR No.04114: Fleshy roots are short (<10 cm) with blunt end and light colour, arranged in wide angles from the axis (diverged type) and with excellent storage quality of Safed musli (Chlorophytum borivilianum) INGR No.06023 : Superior gel rich clone (2191.18 g/ plant) of Aloe INGR No.06024 : Superior aloin-A rich clone (26.13%) of Aloe INGR No.06025 : High starch containing clone (13.32%) of Giloe INGR No.07041 : Compact plant type with high andrographolide content of Kalmegh DWS 327 (INGR 11026): A dwarf (<30 cm) Ashwagandha (Withania somnifera) pure line DWS-6 (INGR 13047) : A new Procumbent type plant of Ashwagandha (Withania somnifera) DGS 22 (INGR 13041): a high fruit producing Madhunashini germplasm. DPO 14 (INGR 11035): An early maturing (85-90 days) and high harvest index Isabgol (Plantago ovata) mutant DWS 296-4 (INGR14010): A golden yellow colour leaf mutant of Isabgol (Plantago ovata Forsk) Elite PGRs registered with NBPGR
  • 23. Conservation In situ Community Conservation Ex situ  National Parks  Protected Areas  Biosphere Reserves  World Heritage Sites (Forest Departments)  Medicinal plants conservation Areas  Medicinal Plants Development Areas In situ Ex situ On- farm On-farm •Seed gene banks •Botanical or Herbal gardens •Home Herbal Gardens •Community herbal gardens •Medicinal plants conservation parks •Cultivation of MAPs •Field gene banks •In vitro repository - Land races - Sacred groves - Folk Varieties (Tribal and Rural families) 23
  • 24. BREEDING OF Withania somnifera
  • 25. Antioxidant Adaptogen Aphrodisiac Liver tonic Anti-inflammatory agen Astringent Treat ulcers Bacterial infection Venom toxins Cancer Astringent is a substance which causes biological tissue to contract or draw together Adaptogens help balance our bodily functions via the neurotransmitters of the brain. (sexual stimulant)
  • 26. DWS 135DWS 132 DWS 132 and DWS 135: Promising ashwagandha lines entered in to IVT-I (2013-14)
  • 27. DWS 327: A dwarf (<30 cm) ashwagandha (Withania somnifera) pure line (INGR 11026) DWS 327 JA 134
  • 28. DWS 327 DWS-6: A new Procumbent type plant of ashwagandha (Withania somnifera) (INGR 13047)
  • 29. First time in ashwagandha male sterile line has been developed
  • 30. Pollen germination in MS stigma Pollen tube growing Pollen tube reaching to ovary No Pollen germination on selfing MS Pollen MS anther
  • 31. Pollinators in male sterile plants Ants House flies Honey Bees 3rd day after pollinationMeiosis was normal in MS
  • 32. Growth study in Ashwagandha Very slow in the initial period up to 75 days Steadily increased from 90 to 150 DAS. The initial dry matter increase – due root and leaves  After 90 days it was due to fruits and leaves.
  • 33.  Young roots had more 12-deoxywithanostramonolide and Withanolide A content per unit root weight  During maturity the content of active ingredients decreased. Chemical changes during growth 12-deoxywithanostramonolide (%) Withanolide A content (%) 0 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0 1 2 3 4 5 6 7 JA-20 JA 134 Linear (JA 134) Linear (JA-20) 90 105 120 135 150 160 0 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0.08 0 1 2 3 4 5 6 7 JA-20 JA 134 Linear (JA 134) Linear (JA-20) 90 105 120 135 150 160
  • 34. Some DUS descriptors identified Dwarf Extra dwarf Trait Descriptor • Plant height • Tall (> 90 cm) • Medium (30- 90 cm) • Dwarf (< 30 cm) • Plant type 1. Erect 2. Semi erect 3. Spreading 4. Prostrate
  • 35. Extra dwarf Trait Descriptor • Branching pattern • No branches • Few (2-3) branches • More branches (4-6) in the bottom • Profuse branches (branch in each node) • Root branching pattern • No branches • Few (1-2) branches • More branches (3-4) • Profuse branches (>5)
  • 36. Variability for plant type Broad leaf Narrow leaf Open type More branching Compact plantErectWhitish leaf
  • 37. Extra dwarf Trait Descriptor • Calyx attachment with berry • Loosely attached • Moderately attached • Tightly attached • Calyx tip • Fully open • Semi open • Closed • Berry (Calyx) shape • Oblong • Roundish • Calyx covering berry • Open • Closed
  • 38. Trait Descriptor • Leaf shape • Ovate • Triangle • Elongate • Leaf margin • Straight • Wavy •Leaf colour • Light green • Dark green •Leaf size • Broad • Narrow •Leaf Curling • Downward curling • Upward curling
  • 39. Variability for leaf characters Moderate wavy More wavy Curved leaf Straight margin Long leaf
  • 40. Some morphologically distinct types Extra dwarf Open type calyx Cluster flowers Less fibrous rootProcumbent type Less flowers More hairy Leaf Colour
  • 41. Variability for berry color Yellow Red Orange Creamy white
  • 43. Jain et al., 2009: J Plant biochemistry and Biotechnology,18(2):249-252 Micro propagation of Withania coagulans Dunal:A critically endangered medicinal plant Induction of shoot bud from nodel explants Proliferation of shoots Root initiation Similar banding pattern of tissue cultured plants
  • 44. MDendrogram constructed using 27 RAPD markers AP 4 Lane: 1 100bp ladder and 2-25 withania accessions
  • 45. 45
  • 46. 84 Isabgol germplasm accessions are being maintained. Collection, purification & maintenance of germplasm-ISABGOL Six Plantago species P. indica, P. arinaria, P. psyllium, P. lanceolata, P. serrana, and P. carnopus
  • 47. Standardize the hybridization procedure No. of floweers emasculated Seed obtain ed % crossing success E & S 203 12 5.91 E & P 212 73 34.43
  • 49. DPO 4 GI 2 DPO 4 DPO 1 DPO 1 TWOVarieties Developed DPO 4 had 30, 15 and 14% higher yield than GI 2, Niharika and local check, respectively DPO 1 had 21, 19 and 14% higher yield than GI 2, Niharika, GI 2, and local check, respectively
  • 50. Early maturing mutant (75-80 d) M2 Flower initiation in 30 days Matures in 80-85 days 100% Flowering in 35-40 days DPO 14:An early maturing (85-90 days) and high harvest index isabgol (Plantago ovata) mutant (INGR 11035)
  • 51. Golden yellow leaf mutant (INGR 14010) DPO 296-4: A golden yellow leaf colour mutant
  • 52. Closed flower mutant Whitish leaf base and stem Short leaf with twisted leaf tip (Dark green) Short leaf with light green colour
  • 53. Short (> 10 cm)leaf with straight leaves Yellowish green leaves Narrow leaf
  • 54. Short (> 8 cm) stem -prostrate Inflorescence within the canopy
  • 57. DPO 185 (resistant) DPO 144, DPO 145, DPO 185 and DPO 333-2 Downey mildew resistance sources identified
  • 58. Physiological study of Isabgol mutants/lines Morning Afternoon Morning Afternoon -25 -20 -15 -10 -5 0 Morning Afternoon Morning Afternoon DPO 18 DPO 22 DPO 30 DPO 38 DPO 44 DPO 61 DPO 69 DPO 70 DPO 72 DPO 78 DPO 113 DPO 126 DTPO 15 DPO 136 DTPO 21 DPO 144 DPO 162 DPO 181 DPO 14 DPO 185 DPO 189 DPO 195 DPO 200 DPO 204 DPO 206 DPO 216 DPO 230 DPO 231 DPO 232 DPO 233 DPO 240 DPO 241 DPO 256 DPO 264 DPO 266 DPO 267 DPO 269 DPO 270 DPO 274 DPO 280 DPO 281 DPO 282 DPO 283 DPO 296 DPO 307 DPO 311 DPO 312 DPO 332 DPO 334 DPO 352 DPO 356 DPO 362 DPO 371 DPO 395 DPO 396 DPO 415 DPO 422 DPO 431 DPO 433 DPO 437 Leaf water potential of Isabgol lines at saturated soil moisture and water deficit conditions. Irrigated Drought
  • 59. Tetraploids were developed in isabgol Out of 17 lines were screened for its ploidy level (diploid vs tetraploid) using FACS analysis and tetraploid has been confirmed in 97 lines File:DTPO4-2op Date:10-03-2011 Time:12:15:18 Particles:1068 Acq.-Time:236 s 0 200 400 600 800 1000 0 20 40 60 80 100 FL4 DAPI counts 0 200 400 600 800 1000 0 20 40 60 80 100 FL4 DAPI counts partec PAS File:Control 1 (DPO 1) Date:10-03-2011 Time:11:26:27 Particles:770 Acq.-Time:349 s 0 200 400 600 800 1000 0 20 40 60 80 100 FL4 DAPI counts 0 200 400 600 800 1000 0 20 40 60 80 100 FL4 DAPI counts partec PAS FACS analysis 2x 4 x 2x File:DTPO2-1 Date:09-02-2011 Time:17:03:48 Particles:1027 Acq.-Time:255 s 0 200 400 600 800 1000 0 20 40 60 80 100 FL4 DAPI counts 0 200 400 600 800 1000 0 20 40 60 80 100 FL4 DAPI counts partec PAS 4x2x Diploid Tetraploid TetraploidDiploid
  • 60.
  • 61. Raw reads from NCBI Quality filteration / trimming of raw reads Prineq-lite Clean high quality reads Denovo transcriptomeAssembly Trinity Assembler AssembledTranscripts Evaluation of Assembly CDS prediction Annotation SSR prediaction GO analysis Blast2GOBlastX MISA Primer3 Primer designing CLC Genomic Workbench InhouseTranscript Workflow ofTranscriptome analysis
  • 62.  The raw data Plantago sample was downloaded from NCBI (http://www.ncbi.nlm.nih.gov/sra/?term=Plantago).  Raw data was filtered using Prineq-lite.  Adapter contamination and reads with low quality value i.e. an average QV less than 20 (QV < 20) were filtered.  The high quality data of Plantago sample was assembled with CLC genomics workbench on default parameters (mismatch cost = 2, Insertion/Deletion cost = 3) for denovo assembly.  A total of 23,586 transcript contigs were obtained for Plantago sample.
  • 63.  The functional annotation was performed using 23,586 transcript contigs of Plantago sample by aligning against NR database of NCBI using BLASTX.  The annotated transcript contigs were mapped on GO database which showed significant similarity with Solanum tuberosum followed by Vitis vinifera, Solanum lycopersicum and so forth.  The distribution of annotated transcript contigs in specific ontology domains indicate 4192 GO terms were assigned to Biological Processes, 2352 GO terms to Molecular Functions and 5309 terms to Cellular Components.
  • 64.
  • 65. 1=100 bp ladder 2=DPO-185 3=DPO-14 4=Club mutant 5=EC 124345 6=EC 427062 7=G-I-3 8=H -I-29 9=DM -4 10=MIB-121 11=DM -1 12=P -79 13=PB-6-3 14=AMB -2 15=DM -2 16=DM -11 17=P -1 18=MALE STERILE 19=DM-6 20= DM-5 31= MIB-124 32=DM-7 33= P-33 34=JI-9 35= PB-10 -4 36= MIB-2 37=UR-188 38=MIB-4 39=MIB-122 40= PS-19 51=A-24 52 H-I-9 53= DM-3 54= RI-88 55= Mutant 56= H-I-6 57= RI-9809 58= RI-157 59= HI-34 60= PB-96 61=100 bp ladder 62= P-44 63= MIB-2 64= PB-6-1 65= RI-49 66= Sel -10 67= MI-4 68=RI-158 69= P-6 70= RI-130 71= DM-10 72= DM-8 73= G-I-1a 74=G-I-2 75=P.Indica 76=P.arineria 77=P.psyllium 78=P.Lanceolata 79= P.Cerina 80=P.Carnopus 21=100 bp ladder 22=AM -5 23=Palampur-3 24= MSB-8 25= J-1B 26=RI-129 27=PS-17 28= RI-89 29= PB-3-1 30=PB-31 41=100 bp ladder 42= MIB-123 43= PB-80 44= Palampur -2 45= RI-149 46= PB-62 47= MIB-125 48= RI-9808 49= RI-153 50= RI-156 Xdapoem21 marker Polymorphism study in different germplasm of Isabgol
  • 68. DGS 22 DGS 22: a high fruit producing Madhunashini germplasm (INGR 13041)
  • 70. DAA 1 DAA 2 DAA 3 DAA 4 DAA 5 Asparagus adscendens
  • 71. DAR 1 DAR 2 DAR 3 DAR 4 DAR 5 Asparagus racemosus
  • 72. Desmodium gangeticum DDG 1 DDG 2 DDG 3 DDG 4
  • 73. Gymnema sylvestre (22 Acc.) DGS 1 DGS 2 DGS 3 DGS 4 DGS 5 DGS 6
  • 74. • Extent of genetic variability • Knowledge of reproductive biology • Information on the genetics of agronomic traits of interest • Information on the genetics of pathways of secondary metabolism. • Breeding strategies • Availability of efficient analytical techniques for quality analysis
  • 75.
  • 76. Disease Species to be targeted Hypertension Guggul (Commiphora weightiii) Sarpagandha (Rauvolfia serpentina) Terminalia species Diabetes Mamejo (Enicostema axillare) Gurmar (Gymnema sylvestere) Cancer Kokum (Garcinia) species Annona Dengue Papaya (Carica papaya) Immunomodulation Ashwagandha (Withania somnifera) Kalmegh (Andrographis paniculata Malaria Artimesia (Artemisia annua) Women health Shatavari (Asparagus racemosus) Asoca (Saraca asoca) Parkinson’s or Alzheimer disease Velvet beans (Mucuna pruriens) Laxative Isabgol (Plantago ovata) Senna (Cassia angustifolia)