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Citrus Breeding Kevin M. Crosby History • Early agriculturalists selected natural hybrids and mutants for seed propagation • 1800’s- grafting and cuttings became popular to propagate best varieties • 1900’s- artificial cross-pollination practiced Seedless fruit of the Tahiti lime More than 70 percent of all citrus fruits grown in the US are varieties developed by the ARS citrus breeding program. Breeding Goals • Rootstock- tree size, stress tolerance • Scion- fruit color, size, shape, flavor, yield • Disease resistance- CTV, Phytophthora, CVC, Alternaria, Scab, Greening, etc. Rootstock • Dwarfing- Poncirus, some mandarins • Compatability- citrus better than Poncirus • Seedling vigor and scion yield • Fruit quality- size, shape, flavor, juice Rootstock Hybrids • ‘Carrizo,’ ‘Troyer’ Citrange- navel orange x Poncirus, very popular in FL, CA • ‘Swingle’ Citrumelo- grapefruit x Poncirus, very popular in Florida, salt intolerant • ‘Sunki’ x ‘Swingle’ tf- semi-dwarfing, salttolerant, possible replacement for SO in TX Carrizo Citrange Swingle Citrumelo Rootstock Fruit Scion • Vigor, yield potential, cold tolerance • Fruit type- mandarin grapefruit, orange • Fruit quality- flavor, size, seediness, appearance, shelf-life Scion Hybrids • ‘Orlando,’ ‘Minneola’ tangelos- ‘Duncan’ grapefruit x ‘Dancy’ tangerine • ‘Page’ mandarin- ‘Minneola’ x ‘Clementine’ • ‘Oro Blanco’ grapefruit triploid- tetraploid pummelo x grapefruit Interspesific hybridisation Disease Resistance • Viruses- CTV, Psorosis, Exocortis • Bacteria- Citrus Variegated Chlorosis, Greening, Canker • Fungi- Alternaria, Scab, Melanose, Phytophthora Breeding Techniques • Cross-pollination- combine genes from different parents in hybrid progeny • Self-pollination- fix genes of interest in one line to stabilize phenotype (inbreeding) • Mutation- natural or induced genotypic modification Cross Pollination • Combine desirable traits from different genotypes/species and exploit heterosis • Swingle- one of earliest to make extensive crosses for rootstock improvement • Many hybrid rootstocks between Poncirus and Citrus- citrange, citrumelo, citrandarin Self-pollination • Natural mechanism for species to maintain genetic uniformity- pummelo, mandarins • Serious inbreeding depression in citrus overcome by apomixis- nucellar embryony • Important for gene inheritance and function studies Seed Structure Mutation • Natural mutations- ‘sports’ of buds or limbs: ‘Ruby Red,’ most orange varieties • Gamma rays- chromosome breaks cause genotypic changes: ‘Star Ruby,’ ‘Rio Red’ • Chemical and t-DNA- interrupt single genes Population Development • Pedigree- all progeny from specific cross carefully evaluated, limited genetic base • Recurrent selection- diverse populations improved by selection and intercrossed • Mass selection- large population evaluated for a few outstanding individuals Pedigree Method • Most citrus varieties developed by this method- few crosses • Relatively few parents (monoembryonic) as females; various males • Each progeny evaluated from each family Recurrent Selection • Each population developed for important traits- good genetic diversity • Crosses between individuals from improved populations evaluated for superior traits • Most productive over long period Mass Selection • Characterized or heterogeneous populations from relatively few crosses screened • Focus mainly on quantitative traits or genes with incomplete penetrance • Labor intensive but rapid improvement Biotechnology • Protoplast fusion of different genotypes • Gene mapping with molecular markersgene cloning. • Genetic transformation with novel genes to modify DNA- Agrobacterium, biolistics Protoplast Fusion • Isolate cell protoplasts from callus or leaf tissue and fuse in vitro to form hybrids • Mostly polyploid plants regenerated from tissue culture- genetic hybrids • Overcome barriers to sexual reproduction Gene Cloning • Mapping genes in DNA with molecular markers- RAPD, RFLP, AFLP, etc. • Chromosome walking- locate DNA markers adjacent to gene of interest, clone gene inside bacterial plasmid • cDNA cloning- isolate genes from mRNA Map-based Cloning 120 kb BAC SCAR CGTTGA- part of FR gene RFLP but no RAPD RFLP RAPD .4 .6 1 CM FINGERBLIGHT RESISTANCE GENE AFLP Genetic Transformation • Insertion of cloned gene sequence into DNA (genome) of desirable plant • Modify single trait while maintaining good attributes of parent- SO with Ctv gene • Insertion point in genome not targeted Texas Priorities • Salt and drought tolerance • CTV and Phytophthora resistance • Cold and heat tolerance • Fresh market fruit- size, sugars, low acid Past Achievements in Texas • ‘Ruby Red Grapefruit’- bud sport of Thompson in LRGV, changed market • ‘Star Ruby’- irradiated seedling of ‘Hudson,’ darkest red grapefruit • ‘Rio Red’- irradiated budwood of ‘Ruby Red,’ most popular red grapefruit today Past Achievements in California • Hybrid mandarins- ‘Kinnow,’ ‘Pixie’ • Triploid seedless grapefruit- ‘Oroblanco’ • Hybrid red pummelo- ‘Chandler’ • Rootstocks- citranges, citrumelos Past Achievements in Florida • Tangelos- ‘Orlando,’ ‘Minneola,’ ‘Page,’ • Grapefruit- ‘Marsh seedless,’ ‘Duncan’ • Tangors- ‘Murcott,’ ‘Temple,’ ‘Fallglo,’ ‘Ambersweet’ Ambersweet, a coldhardy orange variety Shasta Gold, a large, seedless tangerine released 2002 Current Variety Development • New triploid, seedless mandarins- CA, FL • New salt tolerant, dwarf rootstocks- CA,TX • New low acid grapefruits- CA • New fusion product rootstocks- FL Current Molecular Research • CTV resistance gene cloning- CA,TX,FL • Low acid gene mapping- CA • Fruit development gene mapping- CA,FL • Chromosome Isolation and Fusion-TX Future Goals • CTV resistance gene in susceptible scions and rootstocks • Phytopthora resistant, salt tolerant, high yielding rootstocks • Fruit- sweeter, seedless, longer shelf life Breeding Strategy for Texas • Increase effort in transgenics development • Increase emphasis on fruit quality and earliness for fresh market expansion • Increase research into genetic cold tolerance