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Studies on In vitro Propagation of Peony (Paeonia spp.) in Kashmir

KrishiKosh

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Title Studies on In vitro Propagation of Peony (Paeonia spp.) in Kashmir
 
Creator Rather, Zahoor Ahmad
 
Contributor Paul, Dr.T.M.
 
Subject In vitro propagation, herbaceous peony, media browning, tissue culture, tree peony
Floriculture
 
Description PhD Thesis submitted SKUAST Kashmir
Peony is an important ornamental plant used for landscaping and cut flower production. Propagation coefficient of peony is very low through conventional methods. Present studies on in vitro propagation of Peony (Paeonia spp) were conducted for the development of a micropropagation protocol. Investigation was carried out in the Biotechnology Laboratory of the Division of Pomology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar during 2006-2009 using herbaceous peony (Paeonia lactiflora) cv. Sara Bernhardt and tree peony (Paeonia suffruticosa) cv. Black Dragon.
Herbaceous peony (Paeonia lactiflora) cv. Sara Bernhardt
Herbaceous peony (Paeonia lactiflora) cv. Sara Bernhardt is a very excellent cut flower with good vase life. It is highly demanded for its attractive and sweet scented fragrant flowers. Development of a micropropagation protocol is very important for bridging the gap between demand and supply of propagating material of this plant which has a very low multiplication rate in nature. Explant contamination and browning are serious obstacles encountered during in vitro propagation of herbaceous peony. Explant contamination is high as the buds grow under soil. Treatment of different types of explants for 30 minutes in a solution of megapin (0.03%)+ bavistin (0.01%) followed by surface sterilization with mercuric chloride (0.1%) for 10 minutes + 70% ethyl alcohol for 10 seconds proved best sterilant combination giving maximum aseptic cultures in all types of explants. Next best treatment was mercuric chloride (0.1%) for 10 minutes + 70% ethyl alcohol for 10 seconds. Browning is a serious problem with peony. All types of explants resulted in media/explant browning which was controlled by culturing in media supplemented with polyvinyl pyrrolidone (5.0 g/l) and incubation under refrigeration (4 oC) for 48 hours. Antioxidants (ascorbic acid and citric acid) failed to control browning. Among the explant types, maximum media browning was observed with ovary and leaf segments (89.09%) followed by stem segments (54.78%) whereas minimum with terminal buds and petiole segments (45.00%) on media supplemented with ascorbic acid (0.2 g/l) and incubated under light. Explant source and harvesting stage had a significant effect upon browning. Minimum media browning was noted when unexpanded inner leaves of terminal buds were used as explants or explants were collected from forced stock plants.
Explant type, media and growth regulators had a significant influence on callus initiation and growth. Maximum callus initiating cultures (63.17%) and callus fresh weight (321.82 mg) was obtained with BA + 2,4-D (0.50 + 2.00 mg/l). Covered leaf segments gave maximum callus initiating cultures (66.82%), callus fresh weight (281.96 mg) and minimum days to callus initiation (10.404) followed by forced leaf, stem and petiole segments. Highest callus proliferation grade of 3.68 was obtained in ½ MS medium supplemented with BA + 2,4-D + GA3 (0.50 + 1.00 + 1.00 mg/l). Various combinations and concentrations of BAP, NAA, IBA, IAA, 2,4-D and GA3 failed to induce organogenesis in the callus derived from different explants.
Establishment of underground and terminal buds was significantly affected by media and growth regulators. Maximum shoot length and leaf number/shoot was obtained on full strength MS media fortified with BAP + GA3 (0.50 + 0.10 mg/l). Shoot proliferation varied with the explant type recording 6.20 and 2.36 axillary shoots/explant with terminal buds and underground buds on full strength MS media supplemented with BAP (1.00 mg/l) and BAP + Kinetin (0.50 + 0.50 mg/l), respectively. Length of axillary shoots and leaf number/shoot was maximum with BAP + GA3 (0.50 + 0.10 mg/l). Half strength MS medium supplemented with IBA or NAA (0.25, 0.50 & 0.75 mg/l) was used for root initiation. Healthy in vitro shoots were incubated under complete darkness for 15 days in root initiation medium and then transferred to hormone-free root development medium under light conditions. Maximum rooting (46.92%) and maximum root number/shoot (2.62) was recorded on medium supplemented with IBA (0.25 mg/l). Ex vitro survival of the rooted plantlets was 46.67% under greenhouse conditions in pots filled with soil, well decomposed farmyard manure and sand (1:1:1).
Tree peony (Paeonia suffruticosa) cv. Black Dragon
Tree peony (Paeonia suffruticosa) cv. Balck Dragon is an important landscape plant but rare because of very low propagation coefficient. New growing shoots pinkish in color were collected in spring. Shoots were cut into nodal segments and surface sterilized with different sterilant combinations. Maximum aseptic cultures to the tune of 59.01% were obtained when explants were kept immersed in a solution of 0.01% bavistin + 0.03% megapin for 30 minutes and surface sterilized with 0.1% mercuric chloride for 10 minutes + 70% ethyl alcohol for 10 seconds. Survival of the explants ranged between 62.71 to 83.85% but the differences between treatments were statistically non-significant. Nodal segments inoculated in the MS medium resulted in media browning which was maximum in media supplemented with ascorbic acid (26.07%) and in control (22.78%). Polyvinyl pyrrolidone (5.0 g/l) proved best chemical for controlling media/explant browning. There was practically no media/explant browning when explants were inoculated in media supplemented with PVP (5.0 g/l) and incubated under refrigerated conditions at 4oC for 48 hours. Maximum callus initiating cultures (78.81%) and callus fresh weight (512.290 mg) was obtained when in vitro shoots were cultured on medium supplemented with BAP + GA3 (2.00 + 1.50 mg/l). BAP alone or in combination with NAA, IBA, IAA, 2,4-D and GA3 failed to induce organogenesis in callus. Development of shoot buds in callus was observed with different concentrations of TDZ (0.25 – 2.00 mg/l) only. Shoot bud initiating cultures varied non-significantly between 49.39 to 55.44%. Maximum shoot buds/culture (12.80) were obtained on full strength MS medium supplemented with TDZ (2.00 mg/l).
Establishment of nodal segments ranged between 66.14 to 84.21%. Growth regulators significantly affected shoot length and leaf number/shoot which was 0.89 cm and 4.85 with TDZ (1.00 mg/l), 1.46 cm and 9.22 with BAP (1.50 mg/l) and 1.86 cm and 9.86 with BAP + GA3 (1.00 + 1.50 mg/l), respectively. Proliferation via axillary shoots was significantly affected by BAP concentrations recording maximum proliferating cultures (53.93%) and axillary shoots/culture (4.62) with BAP (1.50 mg/l) and minimum of 39.16% and 2.27 with BAP (0.25 mg/l). Adventitious shoot buds developed with different concentrations of TDZ. Non-significant influence was observed on proliferating cultures which ranged between 59.53 to 84.21%. Number of shoot buds/explant significantly varied between 13.45 with TDZ (0.50 mg/l) to 18.20 with TDZ (2.00 mg/l). Satisfactory growth of adventitious shoots was obtained on medium containing BAP (0.25 mg/l). Rooting of in vitro shoots was tried in two media. Shoots were kept in root initiation medium containing IBA (0.1, 0.2, 0.5 mg/l) for 10 days in complete darkness and then transferred to hormone-free root development medium under light. No root initiation was observed even after 2 months of culture in root development medium.
SKUAST Kashmir
 
Date 2016-12-20T13:25:59Z
2016-12-20T13:25:59Z
2010-12-20
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/91453
 
Language en
 
Format application/pdf