‘Therefore, I shall talf,on innovations and applications of Biotechnology in (@ — Production of quality planting material and (i) Genetic improvement of fruit crops 4 f @rduciion of quality planting ‘material tn frutt crops + Meristem culture Getic mparvement of fut crops + Micropropagation i Sex de ao Ov eCenaBSTissue cultural innovations related to meristem culture & micropropagation Haberlandt (1902)"grew palisade cells from leaves of various plants and proposed concept of totipotency i.e. ability of plant cells to differentiate into functional plants when cultured in vitro under controlled conditions Gautheret, Nobecourt and White in 1930's - Callus cultures — cells kept alive — but they did not grow into plants Carrot plant from root cells Tobacco plants from single cells (Stewarde! al, 1964) (Vasil & Hilderorandt 1965) % = WO Mapes, AE Kent ROMOBIEN [Vasi, V_and AC Hidebrandt 143:20-27] nce 150,889.80 [" Habertandt, G. 1902. Prlanzenzsiien. Site-Ber. Mat. Nat. Kl. Kais: Akad. Vilss. Wien 111: 69-92)Meristem culture: A technique for eradicating viruses + Morel and Martin (1952)" developed meristem culture technique for virus eradication in Dahlia + Plant shoots grow by virtue of their apical meristem, * Apical meristem is usually a dome of tissue located at the extreme tip of shoot. it is about 0.1 mm in diameter and 0.25 to 0.30 mm in length. % Vascular bundles are absent in meristem tips and they have high auxin concentrations. Therefore, meristems are generally free from viruses. (1 Morel, G. and ©. Mertin. 1952. Compt Rend. Acad. Sci Ser. D 295:1324-1925)Meristem culture is the only method available to date for _ freeing vegetatively propagated plants from viruses. Once virus free plants are produced through meristem culture, these can be maintained indefinitely in tissue culture and used as [Thom starting material for production of quality planting material. Virus- free starting material Mericlones Ss) Virus TestThermo- + encapsulation-dehydration cryo-therapy of shoo! tips for efficient eradication of ASGV from (A) ASGV infected in vitro shoots without thermatherapy (left) & with thormotherapy at 26 4C (day) and 32 °C nigh (ight B) Died shoot tip after encapsulation dehydration cryotherapy in LN. (C) Callus on 10% surviving shoot tips (0) Direct regeneration of shoots (witout callus) on 80% surviving shoot PN tins ater 4 weeks. 7 > \\, | (©) Receneration of 3 sizes of shoots (LP = Leat Primordia) Eifect of 4 week's thermotherapy [36 °C (day) and 32 °C (right)] on growth of in vitro plants in tissue culture Dera EOC Meenas! No. of shoots/plant (ecru Cao} Thermotherapy 100 25 4.7 (Leaf colour yellowish) Control 100 35 2.5 (Leaf colour green) [Zhao et al 2018, Plant Disease 102:1574-1580]More wirus-freedom is achieved with Thermo- + Shoot tip culture (STC) + Gryo-therapy (Cryo) ASGV eradication (%) 0 0 20 40 Duration of Shoot regeneration thermotherapy ——___ (Weeks) ST oa 0 100 2) 88.9 4 77.8 6 | | | | oo a er © “Heath Pk Longitudinal sections of shoot tps shawing immunolocalization (purple colour) of ASGV. Green circles Indicate virus-free Apical Domes.Four stages of micropropagation” 5 : | stage 0 || Selection and maintenance Micropropagation is an art and |_S'89° || orheaithy mother plants. science of multiplying plants in Excision of explants, sterilization and establishment of in vitro cultures. U Repeated multiplication of propagules (shoots, embryos ete) in vitro. Rooting of shoo's and germination of embryos a Hardening of in vitro plants and their transfer to soil [t Murashige, T1974. Ann. Rew. Plant Physiol 25:135-166)Advantages of micropropagation Production of virus-free plants through meristem culture is possible Enormous rate of multiplication Year-round production of plants In dioecious plants female plants can be selectively multiplied True to type and vigorous plants Uniformity in growth and bearing in the field Higher yield comparedito traditional methods There are about 150 private owned commercial tissue culture units in the country ICAR, CSIR, DBT and SAUs are also involved in the activity Quality standards are well developed, can easily be adopted and harmonized with international standardsMicropropagation methods Method Characteristics* Direct organogenasis ‘Adventitious meristems on explanis (leaf, stem, floral tissues) differentiate into shoots without forming callus. Indirect organogeness Direct somatic embryogenesis Indirect somatic embryogenesis Explants de-differentiate into callus followed by induction of shoot primordia on callus and finally re-differentiation of shoot primordia into shoots. Development of embryos from somatic cells of explants without forming callus. Induction of callus on explants followed by development of embryofembryo lke structures oon the callus Multiple shoot formation trom apicalaxillary buds Initiation and development of multiple shoots from apical/axillary shoot tips. somacional variations. * Methods with green fonts are highly suitable for micropropagation, while methods with ted fonts can only be used for micropropagation if they do not producein Thompson Navel 5 Maximum no. of shoots were produced on MT (Murashige and Tucker, 11963) basal medium supplemented with NAA (0.1 mg/l) and BACT mg!) ss shoots Maximum rooting was recorded on medium supplemented with 0.5 mg/l NAA. Organagenesis in internodal stem cutting (A) and in shoot tip (8 [Esmeina and Dehestan|, 2015. Etherean Biologist Romania) 89-14]Indirect organogenesis in mangosteen | Satwmenssom maim itennig Garcinia mangostana omegesic ca i tl tenn piss sae induction of callus in lear] | Posten Se ene expiants of mangosteen on MS + BAP (2.22 uM) + Thidiezuron — (TDZ) (227 UM), Callusing stars from mid nb (a) Shoot buds on nodular callus (b) Multiple shoot buds [c) Shoot on medium containing 2.22 uM BAP and 2.27 pM TDZ (d) Multiple shoots of mangosteen. [Ocsim et al 2013. Asian Journal of Plant Sciences 12:279-284]Direct somatic embryogenesis in guava (Psidium guajava L.) Development of embryo or embryo like structures from somatic cells without forming callus. Induction and proliferation of multiple somatic embryos from immature zygote embryo on MS +2,4-D (2 mqil) + L-glutamine (400 mal) + Malt extract (500 gi!) Maturation of somatic embryos, plantet germination, rocting, elongation of piantiets, shoot proliferation, plant maturation, hardening and potting of plants [Bajaai etal 2016. Brazilian Archives of Biology & Technology 58:150-1702, 4-D (2 mg/l) was the best concentration for inducing somatic embryogenesis in Psidium guajava L. among all the four genotypes tested in terms cf highest frequency and intensity of embryogenesis AS: Alahabad Safed La: Latt 5d. Saar (L-49) She Shweta CONCLUSION % An effident reprocucibie regeneration somatic embryogenesis protocol was developed in out genotypes cf Psidium guajave L. using immature zygotic embryo as starter expla. Best somatic embryogenesis induction wes obtained on MS basal medium supplemented with 2, 4-D (2 magi, L-glutamine (400 mail), 6% sucrose and Malt extract (500 maf Dajpai etal 2016. Brazilian Archives of Biology & Technology 5950 170]Indirect somatic embryogenesis in (A) Callus @.C) Embryegenie call (0) Stages of somatic embryos (E-F) Sections of ealy toipedo embryos (S) Regenerated plant on % MS + GA (0.05 mgi (H-!) Plants in pot. Strawberry ( Fragaria sp.) fects ot pint orowh requalors on stavbery somatic emanogenesis Gia cca Ga be st se tnerode 60 Te st Geena Peete aetna foamed ens “There wes nol much diference in fel performances ‘of control and embiyo derived pits, Ganon ‘Coot Eesieyo dered plan Moan = SE Mean 2 St Leaf ounberplae 287 48s Narber of umerphat 34z id 59 Number of kuplane 1212 1.2 1062 Average fuk welt (g) 104 LA 107 [Biswas ef al 2007 Plant Cell Tissue & Organ Cara 99-40-54Protocol for producing multiple shoots in Citrus limon and their genetic integrity as analysed by RAPD ‘Shoot initiation ‘rom nodal segment explants of C limon cv. Kaghazi Kelan cultured on (a) growth regulater-ree MS medium; (b) Multiple shoot formation from nodal explan's cultured on MS medium supplemented with BA (1.0 mol) + Kinetin (05 mgf) + IBA (05 maf); (c) Multiple shoot formation from nodal explants cultured on MS medium supplemented with BA (0.25 mg/l) + Kinetin (1.0 mall) + IBA (1.0 mal; (d) Root induction from in vitrosaised shoots on growth reguator-free MS medium, and (e) Aplantlet kept on soil: sand rrixture | Agarose gel image showing the DNA amplification profile generated by primer OPGO7 (a) and OPGOB (b) from eight seedless individuals plants (anes 1-8); M, 100 bp ladder; anc C snormal plant [Goswamiet 2. 2013. Physiology & Molecular Biology ef Plants 49.137:145)Multiple shoot formation in Almond (Purunus amygdalus Batsch) FON in Pe eA 0.005 hol aan al “ 0.025 aM IBA 0.25 LM [BA f “ (BA F- uM IBA Effect of [BA concentrations on induction of multiple shoots in Almond. All above media contained 1 4M BAP and cifferent concentrations of IBA 0 to 5 uM) Media containing 1 uM BAP + 0.005/0.025 4M IBA are most effective for induction of muttple shoots. 1 0.005 1 0.025 0.25 25 o 8 o * With several small buds It has been shown that 1 plant can produce 5 plants in 30 days. Therefore, in a year St? Le. 244 Crore plants can be produced from Hisajima, S. 1982. Agric Biel. Chem. 46: 10micropropagation of grape vine (Vitis vinifera L.) from nodal culture Varieties used in the study Chenin Blanc Ugni Blanc Canonannon Explant sterilization & culture Stem segments were collected from stock plant grown in greenhouse Thoroughly washed with tap water containing ‘Tween 20 Sterilized with Sodium hypochlorite (NaOC}) for different time duration (67 and 9 minutas) Explants (2-3 om long nedes) were cut and cultured on MS medium supplemented with BAP (0.5 mg/l) IKinfo ot al. 2017, Afticen Journal of Biotechnology 16:2083-2001](A) Shoot induction on MS medium supplemented with BAP (0.5 mg/) (B) Shoot multiplication on medium containing 3AP (0.5 mai). (C) Shoot multiplication on BAP (0.5 mg/l) + IBA (0.1 mg) Root induction in MS medium supplemented wih different. IAA concentrations CeCe potted in vitro eon UgniBlenc Chenin Blanc CanonannonCommercial micropropagation of banana by Jain Tissue Culture, Jalgaon (Annual production of about 3.5 Crore banana plantets’ ‘Vrs indexng Disease freedom and age uniformity are major advantages. Conventional planting material: 90% flowering; Harvesting in 17 Intervals (22 months) Tissue cultured planting material: 99% flowering: Harvesting in 5 intervals (12 months)Commercial micropropagation of strawberry at KF Bioplants Pvt Ltd., Pune (Annual production of about 4.0 Crore plantlets) avs Initiation of cutures in vitro (Stage-1) Multiplication of In vitro rooting shoots (Stage-2) (Stage-3) Hardening in Shade/Green nouses Piants ready for packing Plants ready for (Slage-4) delivery3 & 3 E 3 2 2 | 3 2 8 z 8 & g 3 8 g 8 a £ 5 e 5 3 = 8 a > £ o 8 £Advantages of tissue cultured propagules 25% higher yield than the traditional planting material “No mortality in field compared to almost 27-30 % losses in traditional planting material in first few weeks after plantation. “ Healthy & vigorous plant growth resulting in more runners & hence more & better quality fruits.Several plant species are commercially micropropagated in India. Many of them are vegetatively propagated Vegetables (6): Asparagus, Cabbage, Cauliflower, Onion, Potato and Tomato Fruits (12) : Apple, Banana, Chery-apple, Citrus, Date-palm, Grape, Mango, Papaya, Peach, Pineapple, Pomegranate and Strawberry | Omamentals (34); Agalonoma, Alocasia, Alpinia, Alstroomeria, Anthurium, Calathea, Callalily, Camation, Cestrum, Chrysanthemum, Cordyline, Croton, Delphinium, Dendrobium. Dracaena, Eryngium, Ficus, Freesia, Gardenia. Geranium, Gerbera, Gladiolus, Hydrangea, Impatiens, Iris, Lily, Limonium, Maranta, Orchids, Phalaenopsis, Philodendron, Rose, Spathiphyllum and ‘Syngonium Plantation crops (11): Black pepper, Cardamom, Coffee, Ginger, Miscanthus, Rubber, Sugatcane, Tamatind, Tea, Turmeric and Vanilla Forest trees (9): Bamboo, Callistemon, Eucalyptus, Jackfruit, Kedamb, Pinus, Salvadora, Simaruba and TeakEmbryo culture/Embryo rescue Overcoming seed dormancy Shortening breeding cycle To overcome post-fertilization barriers n distant crosses ‘To breed seedless varietiesOvercoming seed dormancy in stone fruits through embryo culture Germination stages. (A) An apricot seed with shell, (B) en apricot seed Isolated from shel; (C) @ whole embryo isolated from shell and testa, (D) the separation of cotyledons from embryo; (E) embryos without cotyledons cullureé on Murashige and Skoog medium: Peach aps A « # Apricot A > # Wild cherry ie B c (A) apricot embryos (wiinout cotyledon) successtuly germinated on MS medium containing 0.5 mg'L~ benzyl aminopurine (BAP) + 3.0 mg L~’ gibberellic acid (GA,); (B) peach embryos (without cotyledon) successfully germinated on MS medium containing 0.5 mg'L-! BAP + 3.0 mg:L~! GA: (C) wild cherry embryos (without cotyledon) successfully germinated on MS medium containing 05 mg" BAP +20 mg-L' GAs [San etal 2014 Hort Science 49.294.296]Shortening the breeding cycle of papaya (Carica papaya L.) by culturing embryos treated with ethrel (@) 45 days old fruts (DOF) treated with Etre! (100, Lilt) for 10 days (10 DT) and 20 days (20 DT) (b) 60 DOF treated with Ethre! for 10 DT and 20 OT (c) 75 DOF ‘reated with Ethrel for 10 DT and 20 DT (a) Embryo germination rate in untreated 105 to 180 DoFs (2) Seedlings irom 69 DOF treated fer 20, ays with water (Control). (o) Seedings from 60 DOFs trealed jor 20 days with Ethrel (100 ul) In Papaya, it takes 6-9 months trom pollination to seedling establishment. Ethrel (100 pill) treatment to immature fruits (45 days onwards) attached to plants for 10-20 days could shorten the breeding cycle by approximately 3 months. [Tamaki ef al. 2011. Plant Cell Tissue & Organ Culture 106:225-233)Embryo rescue in mango crosses (A) In vitro growth of immature hybrid ‘embryo (6) Secondary root incuction In vito plants ready — for transfer/gratting (0) Totapuri root stock (EH) Ex vito shoottip grafting (I) Field establishment (J-L) Flowering and fruting + Zygotic embryos from different crosses were excised aseptically at 5 to 8 weeks post- pollination for in vitro plantlet generation More than 60% embryos formed in vitro shoots + Shootttip grafting was practiced to avoid heavy mortality of in vitro plants when directly transferred to soil [Sahijram et ai. 2013. Joumal of Horticultural Sciences 81-20)Breeding seedless grapes using embryo rescue and MAS ‘Ten female parents and six male parents were used in ths research. Thirteen combinations \were identified based on 2 breeding plan and the characteristics of the cultivars DR1, DR2 DR3, DR6, DR, DRS, DRO, and DRIO ze hybridization descendants of Delight » Ruby Soedloes; FZA2 is a descendant of Flame Seoclese obtained by so-polination. Cos Female chamererisiss Male characteristics Monukta Ssrall seed ace, vnifirw Seon ifira Mornikka Small sae race, wnifers’ Seas, sino + Mena Sana sed race, Viner Sse, # viens Thhenpson Seedles Samal seed ace, 1 vera exes, Haft hanggeo Sexes Sarat eed ase; vine secdoa vnc cxign Thompron Seeds Simallseed ace, Fvuyeru——Stnuspermi vans D9 Thompson Seedlest Sima seed ace, vinifera ———_Stenaspermic, 1 vn 1DR10 «Thompson Sees Small see race, Vung Stenger, 1 ving PAD » Manuka Seeded. ¥ inera Seeds. niena lust Seales ~ Flane Seeds Stenespermi, vf Steomsperic, Fv Blush Seaess* Beichan Stenospernc. 1 iniora Seeded, vino This amurensisbybeid ‘lush Seats ~ Thompson Seodss Stenoxpermic, vf temper viagra Blush Scaless» Hellongian Stenospernic, V vinira Seeded. V anomie [etal 2075 In Vitro Celular & Developmental Biology-Plant 61241248]a {) inforescenes bagged afar palinaton; (@) ovules fermation medium: (8) erry rmedum: (2) plant subeuture ‘an rooting medium) pent in pot covered wah plasie cum: () —bardened "plants in Gigenfouse: —(h) surviving pnts growing in so Tissue oulre radia used n the experiment mano formation medium: Exeisad ovules were cultured on ER medium (Emershad arc Famming 1894) sipplemanted wih IAR (15 magi), GA3 (05 ml), Activated charcoal (0.2%, suerose (60 gf) and agar (7 a) Emono geimnetion medium: After 6-8 weeks, embryos ware excised from the ovules and trarsterrad to germination medium WPM (Loyd and McCown 1980) supplemented wih 8 benzyladerine (0.2 mai), Activated charcoal (1 gl), sucrose (20 ol and agar (7 gl) 2018, Vére Gaiar & Developmen Bltegy Plan S1:247-248)[Days after flowering (DAF) is @ commonly used indiealor for ovule excision tm. + The highest embryo development rale for DRt » Monutka was 40%, and best cutle excision tme was 60 DAF The highest embryo development rate for DRS x Thompson Seediess wat 28 4%, and the best ovule excison time vie 48 DAE. ‘The highest embryo development rals of OR7 » Zhonggue Soedives wa 21%, ‘nd the beat avile excision me wa 48 DAF Ettects of samping tine (DAF) on emoryo recovery [ Days afer | No.of {% Embye development in rose fortlization | owls “oRax | one [oRTs | OR ] DRE (DAF) Jouitured |pacnviea | Tm | ahenagus | x x ssundiess | Seedtess | D1 | ORS we | too | a | sea | sta | a0ab | aie 20 | too | aaa | seen | arab] ea | a0 sz | 100 | sa | a0 | 24a | aebe | 2660 | mbe a | too | ab [20 | 2m | anc | zc | ate ‘Mane wifin 6 Gcumn folowed bythe ware leer are nol sigifcanly Geren! (Ouncaria lle range est P0.05) List sl 2015 fn Viro Celsar ® Developmental Blog Plant 51261-2468]9 naz a LPI amplies 539 bp band only in seedless spe ey oo " Ste ee eee ere re pet ee Linkage cf seedessness in parent vatietes with marker GSLP1. (Ml) Molecular ladder, (1) Blush Seedloss: (2) Monukka; (3) Zhengguo Seediess; (4) Thompson Sesdiess: (5) OR1; (6) DR2: (7) OR3: (8) ORG: (9) DR7; (10) DRB: (11) DRS. (12), R10; (13) F242: (14) Beichun: (15) Hellonaiiana Lietal 2015 in Vitra Celular & Developments! Boiegy Pant St:241-248}]Secdlinss: 2) Monukka: (3) Zherggue Seediess; (4) Thompson Secdlees (6) DRI; (6) DR2. (7) Dra; (@) DRS: (0) DAY, (10) DRS: (1) DAO. (12) DRIO, (13) FLA, (14) Belchur, Conclusions An effecive embry rescue technique was developed in grapes. The best sampling times for ovules of DR1 = Monuicka, DRE » Thompson Seediess. and DRT = Zhengguo Seedloss were 40, 50, and 50 DAF, respectively. GSLP1 fa potontial marker lo detinguish seeded, coadleee, and DR etaine in craze breeding. Eight stains were found te have @ 569-bp band found only in seedless parenis thus, thSome commercial varieties developed using embryo rescue technique Mayfive is a eery-ipening nectiaine (Prunus persica var ‘nucpersics) developed at Horlcultural Crops Research Laboratery, Fresno, Calfomia, from the embryo cultze| program in 1964. Mayfire pens eatller then the earliest ‘commercial culivar, Maybelle. The fruits have desiable| cclour fiemess. and quailty (Breador 0.¥/. Remming) Neclanre Goldcres’ is @ very eary-npenng peach (Prunus persia (L.) Batsch) developed by the Quality Mantenance, Genetics snd Tiansportaion Research Unit of the Herticutural Crops Research Laboratory, Fresno, California. frem the embryo culture program in 1987. ‘Goldereet bears tractive, wel colored, round, yolow-feshed fruit and ls invoduced to provide quality ful, very eery in the season for Calfomia enc areas with similar climate, (Breeders: D.W. Ramming and O. Tannet)Recombinant DNA (rDNA) Technology Molecular markers for early sex determination in papaya Molecular markers to study genetic variability Marker assisted selection (MAS) to accelerate breeding Genetic transformation to introduce novel genes Genome editing to modify gene oxpressionstructure 1D, and C, ‘Crick, 1953. Nalure 174-737-728) 1969, Proc Natl Acad. Sor USA 44: 671-882)Transcription Synthesis of mRNA. i‘ Translation Protein Synthesis o> ge rouvreenine Sip jos during 1980-1967 to deviphor mecharism of DNA ® RNAS Prob,Discovery of restriction endonucleases and DNA ligase to create rDNA molecules [Reston enzymes: Smth, HO ard KW Wicox 1970 hol Bick BY 379-291 ‘and many athe] Digestion of tvo DNA moloculs win same resticten enaye say MBE >= 27 elas =: (a ga2 : Providing Steky Ende « ear tc ii¢rraal 6 ining with DWA Ligases procuce ‘NA Lunase: Reviewed by. Lehman, --@aatre 1974, Science 186;790-797 Dl eTTaA ke — =Molecular Markers Abbreviations AFLP cars EST RAP REMAP AFLP SNP RAPD sts SCAR SSR PCR Molecular makers “Ampliied Fragment Length Pelymorphism Cleaved Amplified Polymorphic Sequences Expressed Sequence Tag Inter-Retrotransposcn Ampiied Polymorphism Retroranepozon-Microzatelite Amplited Polymorphism Restriction Fragnent Langth Polymorphism ‘Single Nucleotide Polymorpnism Random Amplited Polymorphic DNA Sequence Tagged Site ‘Sequence Characterized Ampiied Region ‘Simple Sequence Repeat Polymerase Chain Reaction Inter: Simple Sequence Repeat amificationEarly Sex Determination Using Molecular MarkersEarly sex determination in papaya (Carica papaya L.) & Sox in papaya is controlled by a XY chromosome pair, with two slightly diferent Y chromosomes contioling the development of males (Y) and hermaphrodies (Y") Sox of papaya plants can only be temapnoste ‘vie’ Fert | atecag™ after the plats atah repreritiva matuy, te ater 5-3 \ ron IF the sex of the plants is identified at the seocling stage, then a desired ratio of male to female plants (5% meles to 95% females) can be achieved, thereby saving on the input costSequence Characterized Amplified Region (SCAR) Marker W/11 produces an amplizen of 800 bp in only moles and hermaphrocites. This marker can effectively be used for identifying female seedlings before field planting. [chatuvedi etal. 2014. Acta Bot Crow 79(2) 201-298Molecular Markers for Studying Genetic DiversityMolecular variability in some lranian almond genotypes and related Prunus species 2 almond genotypes (cutivated and wid) were collected from 1 sites in Iran. Iiounenesn eta) 2011. Scita Honveuturee 1211110815Morpholegical cluster analysis of 18 moyphalogcal Waits ‘showed to datine groupe. ‘The first group included retaied Prunus speces (except P cammuns) ‘Al the cultivated genotypes and P. communis were placed In the second group. INihoumaneah ota) 2011, Solna Hontioutures 4244)108-113)“Meiecular analysis revealed « igh degise of seperaon and the & genotypes were grauped | Into cise ab agans 2 elisars wilh merpholagcal characters [itoumaneah et a) 2011, Soins Hontioutires 1244)100-18)Molecular Markers in Marker Assisted Selection (MAS)Marker Assisted Selection Accelerates Crop Breeding shrepmeare: nc sms cae pion Selected” oni (1 he basis of Gover band at ce et tant 2-2 Identificetion af closely inked molecular markers (AFLP, RFLP, SSR. SCAR eic)) ta the desied gone Is preliminary requirement of MAS. ‘Choe itis done, selection of desired segregan's can be cone besed on He: presence ofthis molecular maker, * The accuracy of prediction depends on how cisely the marker is Inked to the ddesred gene? + Two closely linked molecular markers further enhance accuracy of prediction. Since this is selection based on presence of molecular marker, rhysical presence of any sitess(bistic or abiotc) isnot requires + Selection can be done in erly stages and itis very quikIdentification of SCAR markers linked to Roa? anthracnose resistance gene in Strawberry. Reid 210 FS Acpiaton coral agucie 20°AFLP Mates > Converted to SCARSTS-Rea2 817 [Lawetasu-Kier el 2005. Theor & Appl GentConelusions DNA buks from a cross between the resisiant cultivar Gaptoa' and the susceptible cultivar Pajaro’ were screened with 110 EcoRI/Msel AFLP combinations Four AFLP markers were found Inked to the enthracnose resistant gene Read, 4 Two of these markers were converted into SCAR markers (STS- Rea2_417 and STS-Rea_240) and screened in a large segregating ‘population including 178 genotypes, The Read resistance gene was estimated to be 0.6 cM from STS- Rea2_417 end 2.8 oM from STS-ReaZ_240. % Large ecale screening showed that 81.4% and 62.8% of the resistant genotypes were correctly predicted by using $TS-Roa2_417 and STS- Rea2_240, respectively [Loreeteau-tanier eat 200, Theor. & Appl Genetes 11 852-870]Development of SCAR markers linked to powdery mildew (Uncinula necator) resistance in grapevine (Vitis vinifera L. and Vitis sp.) CChremesome 15 ‘ot Regent Mi size standard, GE GfGa-47-42; Vb: Vilard banc sistant individuals, S. susceptible ndviduals ‘without termplate DNA added to the PCR band is the upper one with 780 bp i size © conta} say, The dagnaste [Akhurt ot al 2007. Molsculor Breeding 19:102-111]Apple Breeding: MAS and Beyond “ Mos! varlatios of aopie including those mest outsiandhg In the market came out as chance seedling from open palinaion, Ex. Golden Delclous ( possibly a chance hybrid between “Grimes Goiden” and "Golden Renette © The frequent occurrence of sport mulators occuring naturally In vegetalve propagules also led to selection and release of several “Essentially Derived Varieties (EDV). Long serias of these ED varieties can be easily raced in the Red Delicious, Gala and Fujiapple familias, © Other apole vatiaies came cut from controlled crosses and were created by breeders who ablected parenta wit the desired charactere, crossed them, ane walted for tre results. Apole breeding is very lengthy and uncertain. Therefore Some gevelicists refer Itas a "Gross and Piay” epprcach, ene Et RoyalGala Galaxy —Brookficld Gala Obrogala_—_ Gala Schniga taln. 2016 Fruit Seience 324-29)Genetic Transformation and Gene SilencingTypical gene construct and cloning DNA using a plasmid vectoriene pin (0) Biolistic/Gene gunPost-transcriptional gene silencing through RNAi technology ‘Aotisense gene sequence is ifrosuced in a plan els that preduces antsense RNA which pairs with sense RNA at a paca’ gone Io lorm double srended RNA. Double standod NA is not slable and dsintegrated, thereby, nt allawing the paricaar gone f produce protein ent express s in » + rotested RNAS af icoporated info the: RNA owed sleeing. capt (RISC) Wek fargetamessencer NA pent ts [Preeti 199 nature 394 806-811)Genetic Transformation: A Tool for Quick and Selective Improvement Selectivity A desired trait can be introduced into a desired host without disturbing genetic makeup ofthe host “The technique is not confned to cross fertie plants. Theoretically itis | poseible to identity isolate and tranefer any cesirabie gene trom any living ‘organism to plants and vice versa Quick product development Brings in novel diversity Following 3imocriant discoveries made t possible to isolate and trensfer genes across different plant species. ‘Structure anc function of ONA Restriction endonucleases and ligase Plasmids“HoneySweet” (C5), the First Genetically Engineered Plum Pox Virus (PPV) Resistant Plum (Prunus domestica L.) Cultivar oneySweet" was developed in 1994 ewough genetic tanstormation of hypocotyl | explants using Agrobacterum | tumefaciens EHA 101 carying |the plasmid pGAS82GGIPPV- (CP.33 (Scorza et ol. 1984. Pant Call eporis 1418-22) Following greenhouse tseting using graft and aphid inoculations with the M end D ei ad stains of Plum pox vrus Thee year ote = (FPY), C5 was selected and resinbitns Romana ee 3 HoreySwest [scores eto, 2016, Hortscience82:601-003)Transgenic Papaya Resistant to Papaya Ringspot Virus (PRSV) eee E ce} Development of PRSV Resistant Papaya Transgene used: PRSV Coat Protein gene (prst-co) Transformation method used Bioistec Gene Gun Explants used for transformation: Embryogenie cuitures of “Sunrise, Sunset and Kapoho pepeye culver. PRSV Resistant Tranagenics fh 1882 ‘evelopecin 190 1h 1092, PRSV waa discovered wm the distict of Puna on Hawai island, where 89% of Hawaifs papaya wes grown Within two years, PREV was widespread causing severe damage to the papaya industy in hat ars, A fold tral to test a PRSVresisant “Sunrise” wansgeric papaya stated in 1992 By 1985 some now wansgenie cultvars viz Rahbow" and “SunUp" were devetoped Theto cultivars were commercialized th +908) Rainbow is row widely planted ard has helped to save the papaya industry fom devastation by PRS, Transgeric papaya were also develoned fer otne: counties, such as Thallan, Jamaica, Bazi, and Venezuela, [Gonasives 2004, Agile Farum 736-40)