Evolving Complex Hybrids In OrchidsEvolving Complex Hybrids In Orchids

Name: Vitthal N. BhosaleName: Vitthal N. Bhosale

Roll No. 4603Roll No. 4603

Second Largest familySecond Largest family of flowering plants-10% of angiosperms of flowering plants-10% of angiosperms

State-wise distribution of orchids in India. State-wise distribution of orchids in India. (Source: Singh, D.K. 2001. Orchid diversity (Source: Singh, D.K. 2001. Orchid diversity

in India. in India. In: In: Orchid science & commerce)Orchid science & commerce)

New species being New species being described weeklydescribed weekly

What makes orchid BIG ?

Particulars WorldWorld (Kew Checklist of (Kew Checklist of

Orchids, RBG Kew).Orchids, RBG Kew).

India(Ram pal, et al., 2006,

NRC, Orchids)

No. of Genera No. of Genera 800800 184

No. of Species No. of Species 25,000 (Approx.) 25,000 (Approx.) 1300

Name of the stateName of the state Orchid (Number)Orchid (Number)

GeneraGenera SpeciesSpecies

Arunachal PradeshArunachal Pradesh 130130 600600

Sikkim Sikkim 115115 496496

MeghalayaMeghalaya 9898 352352

AssamAssam 8181 191191

KeralaKerala 7777 230230

Present status of orchids in India

Habitat Species Endemic Extinct / Nearly extinct

Endangered

North Eastern India 675 76 18 34

Eastern Himalaya 730 88 18 105

Western Himalaya 255 10 - 44

Peninsular India 267 13 5 25

Eastern India 130 6 - 5

Andaman & Nicobar group of Islands

117 15 2 2

Central India & Gangetic Plains

60 - - -

(Source: Singh, D.K. 2001. Orchid diversity in India. (Source: Singh, D.K. 2001. Orchid diversity in India. In: In: Orchid science & commerce)Orchid science & commerce)

“Market scenario”

Million value (million US $) of the flowering plants in year 2002 in United States

“In 1862 Darwin showed that - orchid's beauty was not “designed” by God to please humans but honed by natural selection to attract insect cross-pollinators.”

The genus Cypripedium (Lady-slipper Orchids) that Darwin Experimented With.

Darwin’s orchid : “The Origin of Species - by Means of Natural Selection”

Orchid DiversityOrchid Diversity

Floral Diversity in Floral Diversity in PlatantheraPlatanthera

TeagueiaTeagueia spp. spp.

21 new species in 2 years 21 new species in 2 years in 80 sq. milesin 80 sq. miles

Arachnorchis gardneri shows three flowers collected from a single locality at Broke Inlet in Western Australia

Diuris where species frequently exhibit a tremendous range of variation, particularly in flower colour and floral markings

Variation within Calochilus campestris

Hybridisation

Natural hybridisationNatural hybridisation

Evolved from crosses between two speciesEvolved from crosses between two species Interspecific hybrids:Interspecific hybrids:

○ Phalaenopsis intermedia= (P. aphrodie x P. rosea)Phalaenopsis intermedia= (P. aphrodie x P. rosea)- Lindley(1853), - Lindley(1853), Phillipines.Phillipines.

○ Masdevallia splendida =(M. veitchiana x M. barlaeana)Masdevallia splendida =(M. veitchiana x M. barlaeana)

○ Vanda Vanda hybrid (Miss Joaquim)= hybrid (Miss Joaquim)= (V. teres x V. hookeriana) (V. teres x V. hookeriana)

Intergeneric hybrids:Intergeneric hybrids:○ Cattleya guatmalensis= (Epidendrum aurintiacum x Cattleya skinneri)- Cattleya guatmalensis= (Epidendrum aurintiacum x Cattleya skinneri)-

Skinner (1861), Gautemala.Skinner (1861), Gautemala.

Ophrys europe

•Mimicry-Mimicry-PseudocopulationPseudocopulation-Sex drive of certain insects-Sex drive of certain insects-Looks like female wasp and -Looks like female wasp and take advantage.take advantage.

Pollination-RewardsPollination-Rewards

Telipogon sp.

SigmatostalixSigmatostalix sp. sp.

OilsOils (triacyl glycerides) (triacyl glycerides) specialized glandsspecialized glands--elaiophoreselaiophores..

Waxy resin crystals

Maxillaria notylioglossa

Resin rewardResin reward: worker : worker bees for nest constructionbees for nest construction

Cochleanthes anatonaCochleanthes anatona

Sepaline false -Sepaline false -nectarnectar

• Frequency of occurrence of intergeneric hybrids within and between nectar less Frequency of occurrence of intergeneric hybrids within and between nectar less and Nectariferous orchid genera. and Nectariferous orchid genera.

Hybrid parentage Both parents nectar less

Nectariferous and nectar less parents

Both parents nectariferous

No. of species (% of fruit set)

38 (16.2) 53 (44.0) 11 (29.8)

*X2 = 41.6.1 1df, P< 0.001

R- Orchids with no rewards (66 spp.)O+ Orchids with floral rewards other than nectar(7spp.) N+ Orchids with nectar (30 spp.)

Percentage of species

Orchid reward group(Davis and Huxey., 1998)(Davis and Huxey., 1998)

• Pollinator attractiveness increases with distance from flowering orchids

Bob B. M. Wong, et al.,2004

Reactions of Andrena nigroaenea males in behavioral tests in the field.

Attractive ness (R

e sponse o f males p er 3 m

in.) n = 12 16 9 11 12 17

Schiestl et al., 2002

Artificial hybridisationArtificial hybridisation

• Milestones:Milestones:• Mr. John Dominy (1856)- Mr. John Dominy (1856)- Calanthe dominiyi = (C. masuca x C. furcata)Calanthe dominiyi = (C. masuca x C. furcata)

• Mr. William Herbert-Mr. William Herbert- Take crossed capsule to maturity Take crossed capsule to maturity

• Many orchid enthusiasts start working but failedMany orchid enthusiasts start working but failed

• Prof. Knudson (1946)-Prof. Knudson (1946)- gives chemically defined media- Knudson C gives chemically defined media- Knudson C

– Seed germination- main problemSeed germination- main problem

• ““Orchid Hybrid Rush”Orchid Hybrid Rush”

• Nona- generic hybridsNona- generic hybrids

• More than one lac hybrids More than one lac hybrids

• Breeding objectives:Breeding objectives:○ Colour, size and substancesColour, size and substances

○ Round, full-form of sepals and petalsRound, full-form of sepals and petals

○ Length of inflorescence and number of flowers/inflorescenceLength of inflorescence and number of flowers/inflorescence

○ Mode of displayMode of display

○ Blooming periodBlooming period

○ Suit as potted ornamentalsSuit as potted ornamentals

○ Climatic regimesClimatic regimes

○ Resistance to diseasesResistance to diseases

• Pre-visualization and breeder’s eye-Pre-visualization and breeder’s eye-○ Cut flower sprays-Cut flower sprays-

Orientation Orientation Number and size of flowersNumber and size of flowers

○ Individual flowers-Individual flowers- Showiness and sizeShowiness and size

Aspasia epidendroides

Floral morphologyFloral morphology

3 sepals3 sepals2 lateral petals + lip2 lateral petals + lipColumn (fused male Column (fused male

+ female parts)+ female parts)Ovary + pedicelOvary + pedicel

Anther cap

Stigma

Pollinia

• Hybrid:Hybrid:

An offspring derived from the cross between two An offspring derived from the cross between two genetically non-identical individuals, in orchids can be of three genetically non-identical individuals, in orchids can be of three types types (Arditti, 1992)(Arditti, 1992)

1.1. Intra-specific hybridsIntra-specific hybrids2.2. Intra-generic hybridsIntra-generic hybrids3.3. Inter-generic hybridsInter-generic hybrids

• Matter of artMatter of art• Darkside: Heredity-Darkside: Heredity-(Arditti, 1992)(Arditti, 1992)

• Long juvenile phaseLong juvenile phase• Non-endospermic seedsNon-endospermic seeds• Maturity of seedsMaturity of seeds• High seeds per capsuleHigh seeds per capsule

• Time of maturity:

Sr. No. Name of Genera Time taken (in months)

1. Calanthe 4-5

2. Cymbidium 3-6

3. Dendrobium 9-17

4. Paphiopedilum 7-13

5. Phaius 6-9

6. Vanda 5-7

(Hegde, 1984)

In vitro micro-propagation of orchid, Oncidium sp. (Dancing Dolls) by using green pods.

Kalimuthu et al., 2003

Kalimuthu et al., 2003

• Hormonal regulation of flowering in a sympodial orchid hybrid Dendrobium Louisae cv. ‘Dark’ (D. phalenopsis X D. veratrifolium)

Goh., 1978

• Effects of Benzyl Adenine (BA) and Gibberellic Acid (GA₃) on the flowering of Dendrobium Lousae cv. ‘Dark’

Treatment No. of pseudo-

bulbs treated

Total no. of buds initiated

Time to bud initiation

(days)

Total no. of buds

developed to maturity

Average length of

inflorescence (cm)#

Plants flowered

(%)

Control 5 0 - 0 - 0

BA 10 -3M 5 6 9(2), 10 (4)* 6 64.3± 4.8 80

BA 10 -4M 5 4 9(2), 10 (2) 4 64.5 ±2.1 80

GA₃ 10 -3M 5 0 - 0 - 0

GA₃ 10 -4M 5 0 - 0 - 0

BA 10 -3M + GA₃ 10 -4M

5 7 7 (2), 8 (5) 7 66.6 ±0.9 100

* Parenthesis indicates no. of buds initiated # Average length ± s.e.

An axillary shoot which developed first as vegetative shoot but after six nodes changed to productive growth and became an inflorescence . (Scale in cm)

Inflorescence produced by the mature pseudo bulb following treatment with BA (10-3 ) Photograph’s taken 2 months after treatment. Scale in cm

RFLP and inheritance patterns of chloroplast DNA in intergeneric hybrids of Phalaenopsis and Doritis

Chang et al., 2004

Contd…

Contd…

• Chloroplast DNA inheritance in the orchid Anacampestris palustris using single–seed Polymerase Chain Reaction.,

Cafasso et al., 2004

•Minisatellite repeat locus located in Chloroplast tRNALEU intron.

•Internal Transscribed Spacer (ITS 1) primers.

Breeding In Important GeneraBreeding In Important Genera

1. Cattleya: Common species used:

Colour genetics- (Hurst, 1925): Two genes ‘C’ and ‘R’ responsible for colours-CcRr x CcRr

○ Phenotypic ratio- 9:7 (Coloured : Albino)

Name of species Number of hybrids

C. dowiana 435

C. trianaei 392

C. mossiae 315

CR Cr cR Cr

CR CCRR CCRr CcRR CcRr

Cr CCRr CCrr CcRr Ccrr

Cr CCRr CCrr CcRr Ccrr

cr CcRr Ccrr ccRr ccrr

(Humphrey, 1960)

• Most crossed genera to Cattleya- Laelia, Brassovola and Sophronitis

• (Cattleya x Lailea) x (Brassovola x Sophronitis)

Potinara (Fordyce, 1995)

Recent trend : Blue Cattleya ., Graineer (2002)

Source- Laeliocattleya canhamina=(Cattleya mossiae x Laeliocattleya purpurata )

• Laeliocattleya puprpurata var. werkhauseri

Laeliocattleya puprpurata ‘Blue Lyre’

• Cattleya hardyana x Laelia pumilapratens

Laeliocattleya rubens x Sophronitis coccinia

Sophrolaeliocattleya

Selfing

2.Cymbidium

• Species used:

– C. eburneum

– C. gigamteum

– C. lowianum

– C. erythrostylum

– C. devonianum

• Cymbidium alexanderi = (C. eburneo-lianum x C. insigne)., (Keith, 2000)

(Pastel coloured hybrid)

Miniature Cymbidium: (Cymbidium Lois Sander x Cymbidium pumilum )., England (1955).

Cymbidium hybrids

3.Vanda

– Vanda Miss Jaquim = (Vanda hookeriana) x Vanda teres., (Tim, 2001)

– Clones of Miss Jaquim: Atherton, Juliet, Hula Girl, Woodlawn, Douglas.

– Hybrids: Approx. 450

– Blue Vanda:– Vanda coerulea – Vanda rosthschildiana (Mootes, 2001)

– Vanda tasselata x Vanda Mimi Palmer

F1 x Vanda Mimi Palmer

Vanda Arjuna

– Vanda cristata x Vanda tricolor

Vanda Paki (Motes, 2001)

• Genomic classification

– Lee, (1991)- studied genomic constitutions of Aranda cultivars-

– Genomic and meiotic properties of Mokaras, trigeneric hybrids involving Vanda(V), Arachnis (A)and Ascocentrum(As)

Ploidy level Genome

Diploid AV

Triploid AVV

Tetraploid AVVV

Crosses Ploidy level with genomic constitution

Remark

2x (Arachnis x Vanda) x 2x(Ascocentrum x Vanda)

Diploids (2n= 38) 1A + 0.5 (As + V)

Triploids (3n= 57) 2A + 0.5 (As + V)

More vigorous and

horticultural desirable for cut

flowers

2x Mokara x 2x(Ascocentrum x Vanda) Triploids (3n= 57)1A + 0.5 (As + V) + 0.5 (As + V)

Lee et al., 1994

4.Phalaenopsis

• Blue Phalaenopsis:

• Phalaenopsis violacea x Doritis pulcherrima ‘ Indigo Bunting’

Dtps Kenneth Schubert ‘Brother's Purple Lip’ x Doritis pulcherrima

Little Blue Bird

White Phalaenopsis- P. amabilis, P. formosana, P. aphrodite.

Pink Phalaenopsis P. schilleriana, P. sandreana, P. lueddemanniana

White flower with pink lip Phalaenopsis

P. Intermedia (P. aphrodite x P. equestris)

Doritis pulcherrima var. coerula Indigo Bunting, Blueberry, Freed’s Blue.

Phalonopsis violacea var. coerulea Sapphire Doll

5.Dendrobium• Species used- Denrobium phalonopsis

• Hybridisation:• Hybrids of intra-generic- Fertile• Hybrids of inter-generic - (Phalaenanthe x Ceratobium)

(Ceratobium x Eleutheroglossum) (Phalaenanthe x Eleutheroglossum )

sterile

Colchicine treatment

chromosome doubling

Amphidiploids

Fertile plants(Amore & Kamemoto, 1993)

Complex hybrids

Hybrids Parental combinations

Bigeneric hybrids:

Aeridachnis Aerides X Arachnis

Aranda Arachnis X Vanda

Brassocatteleya Brassavola X Catteleya

Laeliocattleya Cattleya X Laelia

Vandaenopsis Phalaenopsis X Vanda

Trigeneric hybrids:

Brasslaeliocattleya Cattleya X Laelia X Brassavola

Holtumara Arachnis X Vanda X Renanthera

Quadrigeneric hybrids:

Iwanagara Brassavola X Catteleya X Diacrium X Laelia

Potinara Brassavola X Catteleya X Laelia X Sophronitis

Pentageneric hybrid:

Hasegawaara Catteleya X Brassavola X Broughtonia X Laelia X Sophronitis

Hexageneric hybrid

(Apacia x Brassia)

(Brrapasia serene x Miltonia Fortaleza)

Forgetara Mexico

(Odontoglossum x Cochilioda)

x Odontioda Isabella)

(Miltonia x Oncidium)

(Miltonidium Azetac Gold

Burrageara Sambu River

Brilliandera Gary

Gary., 1982

Conclusion

• Absence of the any crossing barriers makes orchid hybridisation most easier

• Evolution of new species of the orchids is so fast, which provides new genotypes for crossing and getting novelties

• Juvenile phase of orchids can be reduced by using tissue culture facilities, growth hormones

Future Thrust

• Work on systematic breeding approach

• Conservation of highly valuable and endanger orchid species

• Use of biotechnology tools (Molecular markers for varietal identification, Genetic engineering to introduce novel genes, Biological bioreactor for multiplication)

• Standardization and evaluation of most suitable Indian species for hybridisation