Biotechnological approaches of Tuber and Leafy Vegetables ppt.pptx

This ppt mainly focuses on the biotechnological approaches applied to vegetable crops for their improvement.

1. Biotechnological approaches Tuber and Leafy Vegetables Presented by Nidhi Priya Ph.D I Year

2. Broad spectrum of techniques used for altering living organisms for human use. Offers many opportunities for agriculture and provides the means to address many of the constraints placed to productivity. Uses the conceptual framework and technical approaches of molecular biology and plant cell culture systems. Commercial area planted to transgenic crops is in excess of 90 million hectares  (P. Parvatha Reddy, 2012). What is a Biotechnological approach?

3. Conventional breeding programmes are slow and less efficient Inserting new traits Disease resistance Rapid multiplication of plants ( eg . Tissue culture techniques) To produce virus free plants Why do we need to Biotechnological approach in vegetable crops

4. Why Biotechnological approaches are required BIOTECHNOLOGY

5. Major root tuber and leafy vegetables

6. Sweet potatoes are a versatile and nutritious root vegetable crop. Family: Convolvulaceae Origin : Central America Chromosome no. 2n=6x=90 Ancestor - Ipomea trifida Inflorescence - Cyme Moderately drought tolerant crop Optimum temperature: >20 C Ideal temperature for tuber formation is 20-30 C “ Superfood ” by the ‘Centre for Science in the Public Interest’ (CSPI), USA Sweet potato ( Ipomea batatas )

7. The majority of Sweet potato breeding initiatives have as their primary goals as Maintenance or enhancement of tuber colour Shape Quality Yield Improvement of particular processing qualities and resistance to pests and diseases. Biotechnological approaches in Sweet Potato

8. Biotechnological approaches Features Plant tissue culture large-scale production of high-quality and disease-free plants in a short time period. (Butt et al. 2015; Mukhopadhyay et al. 2016) In vitro plant regeneration through meristematic organogenesis of sweet potato Ipomoea batatas L. cv. Owairka Explant used : Apical buds Simultaneously shoot proliferation and rooting, plantlet regeneration. - Elliott 1969 Ipomoea batatas L. cv. Jewel Explant used: Axillary buds Simultaneously shoot proliferation and rooting, plantlet regeneration Contd … Reference: Elliott,1969 and Alconero et al. 1975

9. Contd… Biotechnological approaches Features In vitro plant regeneration through meristematic organogenesis of sweet potato Ipomoea batatas L. var. Gouri Explant : Nodal explant Shoot proliferation, rooting and plantlet regeneration Ipomoea batatas L cv. Purple Explant : Shoot tip Shoot proliferation, rooting, and plantlet formation Reference : Shaji et al ., (2019) and Yang 2010

10. Biotechnological approaches in Cassava Botanically it is Manihot esculenta. Family : Euphorbiaceae Perennial woody shrub Edible parts : Roots Cassava starch plays a very crucial role in food industries. Because of its high The drought tolerance and low demand for nutrients, it can produce acceptable yields even under marginal environmental conditions (Cock, 1982).

11. Biotechnological approaches in Cassava Reference: Stupak et al .,(2006) Biotechnological approaches to cassava protein improvement, Trends in Food Science & Technology

12. LIMITATIONS

13. Biotechnological approaches in Cassava Reference: Stupak et al .,(2006) Biotechnological approaches to cassava protein improvement, Trends in Food Science & Technology Artificial storage protein (ASP1) has been shown to be expressed in Escherichia coli and tobacco at high levels. Embryogenic suspensions of cassava via Agrobacterium-mediated gene transfer was done. Expression of ASP1 gene in cassava (Zhang, Jaynes, Potrykus, Gruissem , & Puonti-Kaerlas , 2003), in which expression levels were higher in leaves than in primary roots was reported. Transgenic approach

14. Contd … RNAi: T his is a type of post-transcriptional gene silencing that controls gene expression in a variety of ways.    RNAi tools were used in controlling whitefly, vectors of cassava mosaic disease (CMD), and cassava brown streak disease (CBSD). It targets the inhibition of V-ATPase A, an enzyme that links the energy of ATP hydrolysis to proton transport across intracellular and plasma membranes of eukaryotic cells Replacement of miR159 (a miRNA targeting the CBSV) precursor with artificial miRNAs from cassava brown streak viruses triggered a higher level of disease resistance in transgenic cassava.

15. Reference: Walsh et al., (2019) Otun e t al., (2022)

16. Tilling: This is a reverse genetic method. Used for detection induced mutations in a specific location. This method has been used to find alleles or characterize gene function in cassava. Use of tilling to detect nucleotide polymorphisms

17. Biotechnological approaches in Leafy Vegetables Name Botanical Name Family Chromosome no. 1. Amaranthus Amaranthus spp. Amaranthaceae 2n=32 2. Spinach Spinacea oleracea Chenopodiaceae 2n=12 3. Palak Beta vulgaris var bengalensis Chenopodiaceae 2n=2x=18

18. Biotechnological approaches in Leafy Vegetables Genetic material Technology Type Purpose Amaranthus hypochondriacus Agrobacterium mediated transformation Agrobacterium tumefaciens strains used: C58- pTiC58 and A281(pGA471) Tissue-specific and light-inducible expression directed by a pea chlorophyll a/b-binding protein promoter in transgenic amaranth plants and their progeny Amaranthus hypochondriacus Agrobacteriummediated transformation Agrobacterium rhizogenes were used: R1000, K599 and BVG strain Protocol for plant regeneration via somatic embryo germination from grain amaranth transgenic hairy roots Amaranthus hypochondriacus Molecular markers SNP markers Genetic diversity and fingerprinting Amaranthus cruentus Molecular markers Linkage mapping QTL mapping

19. Major genetic and genomic resources available for spinach. DNA markers 12 nuclear SSR markers were identified and amplified in spinach Genetic map 110 markers (101 AFLP and 9 SSR markers) were assigned to linkage groups. The gender determination gene was mapped to the linkage group 3 at 101.5 cM nearby five other markers between 97.4−102.6 cM. Bacterial artificial chromosome (BAC) library construction BAC library was constructed from a near-isogenic line (NIL1). Fourteen sequences analogous to known plant disease resistance genes were identified. Of the 14, nine contained nucleotide binding sites leucine-rich-repeat (NBS-LRR) domains. One hundred SSR loci were identified. One primers pair designed from these putative resistance genes was closely linked to the RPF1 locus and cosegregated with the DM1 marker.

20. Limitations of Biotechnological approaches

21. References Yang X (2010) Rapid production of virus free plantlets by shoot tip culture in vitro of purple coloured sweet potato ( Ipomoea batatas L. Lam). Pak J Bot 42 (3):2069–2075. Shaji A, Hegde V, Sailekshmi N, Koundinya AVV (2019) An efcient micropropagation protocol for nutritionally rich varieties of sweet potato ( Ipomoea batatas L.). J Root Crop 45(2):12–18 Elliott RF (1969) Growth of excised meristem-tips of kumura , Ipomoea batatas Linn. poir in axenic culture. N Z J Bot 7:158–166 Alconero R, Santiago AG, Morales F, Rodriguez F (1975) Meristem tip culture and virus indexing of sweet potatoes. Phytopathology 65:769–772 Behera , S., Chauhan , V. B. S., Pati , K., Bansode , V., Nedunchezhiyan , M., Verma , A. K., ... & Naik , S. K. (2022). Biology and biotechnological aspect of sweet potato ( Ipomoea batatas L.): A commercially important tuber crop.  Planta ,  256 (2), 40. Cock, J. H. (1982). Cassava da basic energy-source in the tropics . Science , 218, 755e762 Walsh HA, Vanderschuren H, Taylor S, et al. RNA silencing of South African cassava mosaic virus in transgenic cassava expressing AC1/AC4 hp- RNA induces tolerance. Biotechnol Rep. 2019;24:e00383.

22. References Lightfoot DJ, Jarvis DE, Ramaraj T, Lee R, Jellen EN, Maughan PJ, et al.. Single-molecule sequencing and Hi-C-based proximity guided assembly of amaranth ( Amaranthus hypochondriacus ) chromosomes provide insights into genome evolution. BMC Biol. (2017) 15:74. 10.1186/s12915-017-0412-4. Jamalluddin N, Massawe FJ, Mayes S, Ho WK, Symonds RC. Genetic diversity analysis and marker-trait associations in  Amaranthus  species.  PLoS ONE.  (2022) 17:0267752. 10.1371/journal.pone.0267752. Feng C, Bluhm BH, Correll JC. 2015. Construction of a spinach bacterial artificial chromosome (BAC) library as a resource for gene identification and marker development. Plant Molecular Biology Reporter 33:1996−2005