Callus Induction and Shoot Regeneration of Tomato (Solanum lycopersicum L. cv. Campari) through Anther Culture

Prakash Babu Adhikari,Binod Prasad Luitel,Won Hee Kang 한국원예학회 2011 한국원예학회 학술발표요지 Vol.2011 No.10

In Vitro Callus Induction and Organogenesis in Seedling Explants of Tomato (Lycopersicon esculentum Mill.)

Prakash Babu Adhikari,Binod Prasad Luitel,Doo Won Lee,Cheol Soo Yoon,Won Hee Kang 한국원예학회 2012 한국원예학회 학술발표요지 Vol.2012 No.5

Association of Floral Bud and Anther Size with Microspore Developmental Stage in Campari Tomato

Prakash Babu Adhikari,Won Hee Kang 한국원예학회 2017 원예과학기술지 Vol.35 No.5

Successful anther/microspore culture largely depends on the use of microspores at appropriate developmental stages at the time of culture, which can be specific for each plant species and genotype. This study was carried out to determine the correlation between morphological characteristics, namely floral bud (FB) length and width and anther length, and specific microspore developmental stages in Campari tomato. FBs ranging from 3.55 mm to 18.38 mm were collected, for which FB diameter and anther length were quantified. Furthermore, the developmental stage of microspores contained within these FBs were observed and recorded. The study revealed a strong positive correlation between each size parameters and the progression of microspore development. Moreover, the strongest correlation was observed between anther length and microspore development, indicating that this parameter was more suitable to predict the microspore developmental stage than the FB size parameters. The study also showed wide and overlapping ranges of FB lengths/diameters and anther lengths for specific microspore developmental stages. Subsequent histological observation revealed that this was the result of microspores at multiple developmental stages within a single anther. Moreover, microspores located in the middle of anthers were at a more advanced developmental stage compared to that of microspores at either anther extremity. The results of this study can be readily applied in the collection of Campari tomato FBs/anthers that harbor microspores at particular developmental stages for anther/microspore culture or for further tissue–specific developmental studies. Also, the technique described here can be applied for similar applications in other plant species and genotypes.

Morphological Characterization of Anther Derived Plants in Minipaprika (Capsicum annuum L.)

Binod Prasad Luitel,Prakash Babu Adhikari,Surendra Lal Shrestha,Won Hee Kang 한국육종학회 2012 한국육종학회지 Vol.44 No.4

This study was done to characterize the anther derived regenerants (R1) including haploids and spontaneous diploids of minipaprika (cvs.‘Vine sweet-red’ ‘Vine sweet-yellow’ and ‘Vine sweet-orange’) in glasshouse. Eleven haploids (three, seven and one from red, yellow and orange, respectively) and sixteen spontaneous diploids (five, nine and two from red, yellow and orange, respectively) were grown in plastic pot with three (red, yellow and orange) anther donor (R0) F1minipaprika varieties. Regenerants were characterized for their plant and fruit characters as well as their fruit color and shape. The homozygosity of spontaneous diploid plants of each population was assessed using simple-sequence repeat (SSR) marker analysis. Haploid plants were characterized by reduced plant height, small leaves, short petiole and internode and small flower bud and all haploids showed the sterility and vice-versa in spontaneous diploid lines. The fruit biometrical traits exhibited greater variation within the spontaneous diploid plants and average value of quantitative traits is lower than standard varieties. MR-4 gave the highest yield (150.5 g) per plant followed by MY-6 (140.0 g) and MY-8 (130.5 g) and the lowest in MY-5 (31.5 g). Morphological marker such as fruit color further determinedthe microspore origin of androgenic diploids obtained in anther culture of ‘Vine sweet-red’. Of the fifteen spontaneous diploid plants, fourteen plants were identified as doubled haploids using microsatellite markers (SSR), and these homozygous lines are recommended to use in minipaprika breeding program.

Yield and Fruit Quality of Tomato (Lycopersicon esculentum Mill.) Cultivars Established at Different Planting Bed Size and Growing Substrates

Binod Prasad Luitel,Prakash Babu Adhikari,Cheol Soo Yoon,Won Hee Kang 한국원예학회 2012 한국원예학회 학술발표요지 Vol.2012 No.5

Yield and Fruit Quality of Tomato (Lycopersicon esculentum Mill.) Cultivars Established at Different Planting Bed Size and Growing Substrates

Binod Prasad Luitel,Prakash Babu Adhikari,윤철수,강원희 한국원예학회 2012 Horticulture, Environment, and Biotechnology Vol.53 No.2

This study was carried out to investigate the yield and fruit quality of tomato (Lycopersicon esculentum Mill.)cultivars using different plant bed width and growing substrates in Hwacheon Farm, 2011. Transplants of tomato cultivars ‘Campari’, ‘Temptation’, ‘Annamay’, and ‘Adoration’ were planted at 20 cm, 40 cm, 60 cm, and 80 cm bed width and same cultivars were grown at cocopeat, rockwool and masato in early July, 2011 in a greenhouse. Planting bed size significantly affected the fruit number, marketable fruit number (%) and weight (g), and total yield (g) per plant. Plants grown at 60 cm bed width yielded the highest fruit number (22.4), marketable fruit number (56.0%), marketable fruit weight (356.8 g) and total yield (564.5 g) per plant. Bed width had no significant effect on fruit size and quality characters. Plants grown in cocopeat produced the highest marketable fruit number (56.2%) per plant and yielded the greatest (445.6g) marketable yield per plant. Plants grown in cocopeat substrate produced higher fruit number (5.2%) and total yield (0.7%) than that of rockwool substrate. Fruit size and fruit quality characters showed no significant differences within growing substrates. Hence, planting the tomato cultivars in single row at 60 cm bed width is better approach to optimize the production space in greenhouse and cocopeat is recommended as potential growing substrates for tomato cultivation to increase its yield and fruit quality characters. This study was carried out to investigate the yield and fruit quality of tomato (Lycopersicon esculentum Mill.)cultivars using different plant bed width and growing substrates in Hwacheon Farm, 2011. Transplants of tomato cultivars ‘Campari’, ‘Temptation’, ‘Annamay’, and ‘Adoration’ were planted at 20 cm, 40 cm, 60 cm, and 80 cm bed width and same cultivars were grown at cocopeat, rockwool and masato in early July, 2011 in a greenhouse. Planting bed size significantly affected the fruit number, marketable fruit number (%) and weight (g), and total yield (g) per plant. Plants grown at 60 cm bed width yielded the highest fruit number (22.4), marketable fruit number (56.0%), marketable fruit weight (356.8 g) and total yield (564.5 g) per plant. Bed width had no significant effect on fruit size and quality characters. Plants grown in cocopeat produced the highest marketable fruit number (56.2%) per plant and yielded the greatest (445.6g) marketable yield per plant. Plants grown in cocopeat substrate produced higher fruit number (5.2%) and total yield (0.7%) than that of rockwool substrate. Fruit size and fruit quality characters showed no significant differences within growing substrates. Hence, planting the tomato cultivars in single row at 60 cm bed width is better approach to optimize the production space in greenhouse and cocopeat is recommended as potential growing substrates for tomato cultivation to increase its yield and fruit quality characters.

High frequency somatic embryogenesis and plant regeneration of interspecific ginseng hybrid between Panax ginseng and Panax quinquefolius

Kim, Jong Youn,Adhikari, Prakash Babu,Ahn, Chang Ho,Kim, Dong Hwi,Kim, Young Chang,Han, Jung Yeon,Kondeti, Subramanyam,Choi, Yong Eui The Korean Society of Ginseng 2019 Journal of Ginseng Research Vol.43 No.1

Background: Interspecific ginseng hybrid, Panax ginseng ${\times}$ Panax quenquifolius (Pgq) has vigorous growth and produces larger roots than its parents. However, F1 progenies are complete male sterile. Plant tissue culture technology can circumvent the issue and propagate the hybrid. Methods: Murashige and Skoog (MS) medium with different concentrations (0, 2, 4, and 6 mg/L) of 2,4-dichlorophenoxyacetic acid (2,4-D) was used for callus induction and somatic embryogenesis (SE). The embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 Schenk and Hildebrandt (SH) medium. The developed taproots with dormant buds were treated with $GA_3$ to break the bud dormancy, and transferred to soil. Hybrid Pgq plants were verified by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analyses and by LC-IT-TOF-MS. Results: We conducted a comparative study of somatic embryogenesis (SE) in Pgq and its parents, and attempted to establish the soil transfer of in vitro propagated Pgq tap roots. The Pgq explants showed higher rate of embryogenesis (~56% at 2 mg/L 2,4-D concentration) as well as higher number of embryos per explants (~7 at the same 2,4-D concentration) compared to its either parents. The germinated embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 SH medium to support the continued growth and kept until nutrient depletion induced senescence (NuDIS) of leaf defoliation occurred (4 months). By that time, thickened tap roots with well-developed lateral roots and dormant buds were obtained. All Pgq tap roots pretreated with 20 mg/L $GA_3$ for at least a week produced new shoots after soil transfer. We selected the discriminatory RAPD and ISSR markers to find the interspecific ginseng hybrid among its parents. The $F_1$ hybrid (Pgq) contained species specific 2 ginsenosides (ginsenoside Rf in P. ginseng and pseudoginsenosides $F_{11}$ in P. quinquefolius), and higher amount of other ginsenosides than its parents. Conclusion: Micropropagation of interspecific hybrid ginseng can give an opportunity for continuous production of plants.

High frequency somatic embryogenesis and plant regeneration of interspecific ginseng hybrid between Panax ginseng and Panax quinquefolius

Jong Youn Kim,Prakash Babu Adhikari,안창호,Dong-Hwi Kim,Young Chang Kim,Jung Yeon Han,Subramanyam Kondeti,Yong Eui Choi 고려인삼학회 2019 Journal of Ginseng Research Vol.43 No.1

Background: Interspecific ginseng hybrid, Panax ginseng Panax quenquifolius (Pgq) has vigorous growth and produces larger roots than its parents. However, F1 progenies are complete male sterile. Plant tissue culture technology can circumvent the issue and propagate the hybrid. Methods: Murashige and Skoog (MS) medium with different concentrations (0, 2, 4, and 6 mg/L) of 2,4- dichlorophenoxyacetic acid (2,4-D) was used for callus induction and somatic embryogenesis (SE). The embryos, after culturing on GA3 supplemented medium, were transferred to hormone free 1/2 Schenk and Hildebrandt (SH) medium. The developed taproots with dormant buds were treated with GA3 to break the bud dormancy, and transferred to soil. Hybrid Pgq plants were verified by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analyses and by LC-IT-TOF-MS. Results: We conducted a comparative study of somatic embryogenesis (SE) in Pgq and its parents, and attempted to establish the soil transfer of in vitro propagated Pgq tap roots. The Pgq explants showed higher rate of embryogenesis (~56% at 2 mg/L 2,4-D concentration) as well as higher number of embryos per explants (~7 at the same 2,4-D concentration) compared to its either parents. The germinated embryos, after culturing on GA3 supplemented medium, were transferred to hormone free 1/2 SH medium to support the continued growth and kept until nutrient depletion induced senescence (NuDIS) of leaf defoliation occurred (4 months). By that time, thickened tap roots with well-developed lateral roots and dormant buds were obtained. All Pgq tap roots pretreated with 20 mg/L GA3 for at least a week produced new shoots after soil transfer. We selected the discriminatory RAPD and ISSR markers to find the interspecific ginseng hybrid among its parents. The F1 hybrid (Pgq) contained species specific 2 ginsenosides (ginsenoside Rf in P. ginseng and pseudoginsenosides F11 in P. quinquefolius), and higher amount of other ginsenosides than its parents. Conclusion: Micropropagation of interspecific hybrid ginseng can give an opportunity for continuous production of plants.

Androgenic Response to Different Forms of Minipaprika (Capsicum annuum L.)

Binod Prasad Luitel,Prakash Babu Adhikari,Won Hee Kang 한국원예학회 2011 한국원예학회 학술발표요지 Vol.2011 No.10

Variation in Quantitative and Qualitative Characters in F<SUB>1</SUB> and F<SUB>2</SUB> Populations of Commercial Tomato Cultivar ‘Campari’

Won Hee Kang,Binod Prasad Luitel,Prakash Babu Adhikari 한국원예학회 2011 한국원예학회 학술발표요지 Vol.2011 No.10