Combinations Of 6-benzylaminopurine Research Articles

Micropropagation of Pomegranate (Punica granatum L.) ‘Ganesh’ cultivar from nodal explants.

The present study was carried out to develop a rapid in vitro multiplication protocol using nodal explants. Complete plantlets have been successfully regenerated from node explants. The explants were incubated on Murashige and Skoog (MS) culture medium supplemented with different combinations of 6-benzyl amino purine (BAP) Indole acetic acid (IAA) and kinetin. Multiplication of node explants was achieved on BAP while from other combinations of growth hormone,. The response of various growth regulators was investigated. The combination of       showed the highest rate of shoot induction (100%) and a shoot regenerated about 6 shoots per explants when cultured on MS media supplemented with BAP (4 µM) within 2 weeks. The multiple elongated shoots were obtained on MS basal medium combination of Activated charcoal and growth hormones. Well-developed roots were achieved on MS medium in combination with BAP and IAA as well as 80% of plantlets were survived in the soil successfully in the hardening process. A robust and optimized protocol will be helpful for the mass production of this economical important  fruit .

Effect of Phytohormones on Shoot Proliferation of Doubled Haploid Lines of African Marigold (Tagetes erecta L.)

In vitro propagation of marigold has played a very important role in rapid multiplication of cultivars with desirable traits and production of healthy and disease-free plants. Shoot proliferation is a very important phase of micropropagation. This paper describes an efficient protocol for In vitro micropropagation of Doubled haploid lines of marigold (Tagetes erecta L.) using nodal explants. Murashige and Skoog medium (MS) supplemented with various concentrations and combinations of Benzylaminopurine (BAP), Naphthylacetic acid (NAA), Gibberellic acid (GA3) and Thidiazuron (TDZ) were employed for shoot regeneration in this study, maximum percent sprouting (100%) and minimum days to sprout break (1.48) were found in MS media supplemented with BAP 2mg/L + NAA 0.05mg/L + GA3 1mg/L, while maximum number of sprouts (1.73) and maximum average shoot length (3.98cm) were observed in case of media supplementation with BAP 1mg/L + NAA 0.05mg/L + GA3 0.5mg/L. However, in case of Thidiazuron (TDZ) minimum days taken for shoot proliferation (2.66) and maximum number of shoots per micro shoot (7.04) were observed in media supplemented with BAP 1mg/L + NAA 0.1mg/L + TDZ  0.5mg/L, while maximum length of shoots (4.2cm) was observed in control devoid of any hormones.

In vitro micropropagation of Aloe elegans Tod. using offshoot cuttings

ObjectiveAloe elegans Tod. is an ecologically, environmentally, medicinally, and commercially useful aloe species in Ethiopia and Eritrea. Unfortunately, it is highly threatened due to industrial and urban expansion and traditional mining and agricultural activities. As a consequence, it is included in the IUCN List of Threatened Species since 2013. The plant is getting thinly populated in many parts of the Tigrai floristic region since it is being exploited for traditional and commercial purposes. Therefore, this study was aimed to develop a reproducible, large-scale micropropagation protocol using offshoot cuttings in Murashige and Skoog (MS) media enriched with plant growth regulators (PGRs).ResultsSterilized explants cultured in full-strength MS media enriched with 0.25 mg/L benzyl amino purine (BAP) and 0.10 mg/L naphthaleneacetic acid (NAA) resulted in 100% healthy and live (i.e., initiated) explants after four weeks of initiation study. Unsupplemented initiation media (control) yielded only 14.3% initiated explants. The initiated explants were tested for their shooting response to produce microshoots by incubating in different concentrations and combinations of BAP and NAA for four weeks. Fewer days to shooting (13.0 ± 1.0 days), higher mean shoot number (5.0 ± 1.0), and higher mean shoot length (9.20 ± 2.35 cm) were observed with 1.0/0.50, 1.0/0.25, and 1.0 /0.50 mg/L BAP/NAA combinations, respectively. The rooting responses of the microshoots toward producing plantlets were also tested by incubating them in half-strength MS media enriched with different concentrations of NAA and indole-3-butyric acid (IBA) for four weeks. Fewer mean days to rooting (12.0 ± 1.0 days), higher mean root number (8.0 ± 4.0), and higher mean root length (7.53 ± 3.03 cm) were observed in MS media enriched with 0.75, 0.75, and 1.25 mg/L IBA, respectively. The responses of A. elegans plantlets to primary (in greenhouse) and secondary (in nursery shade and direct sunlight) acclimatization in coco peat, composted soil, and manured soil media were high – with survival percentages of 87.5–97.8% in three to four weeks.

The Effect of IAA and BAP on Root Induction of Cattleya Orchids

Orchid is an ornamental plant commodity that has an important meaning in international trade. Until now, orchids are still the center of attention of farmers and lovers of ornamental plants, because of their bright potential as cut flowers and potted plants. The popular and widespread type of orchid is Cattleya sp. The Cattleya orchid is also called the queen of orchids because of its colorful flowers. Cattleya sp. orchids take a long time to propagate by seed, about 4 to 7 years, so other methods are needed to deal with them. Tissue culture is a method for isolating plant parts such as cells, tissues or organs, and cultivating them in an aseptic environment. The purpose of this study was to determine the optimal interaction of growth regulators indole acetic acid (IAA), benzyl amino purine (BAP), and their interactions with the root induction of Cattleya sp. Cytokinins interact with auxin in determining the direction of cell differentiation. In this research method, differences in concentrations of indole acetic acid (IAA: 0, 2, 4, 6 ppm) and concentrations of benzyl amino purine (BAP: 0, 0, 5, 1 ppm) were carried out. Parameters observed were the number of roots and root length. Data were analyzed descriptively and inferentially using ANOVA. The highest number of roots was obtained in the treatment medium containing 0.5 ppm BAP and 6 ppm IAA, which was 6.33. The highest root length was found in the B0I4 treatment (BAP 0 + IAA 4 ppm). The results of the analysis of the interaction variance between the IAA treatment and the BAP treatment were 0.005, meaning that there was a significant effect of the combination of IAA and BAP on the leaf length of Cattleya sp.

The Effect of Benzyl Amino Purine and 2,4-Dichlorophenoxyacetic Acid on in Vitro Propagation of Pranajiwa (Euchresta horsfieldii (Lesch) Benn.)

Pranajiwa (Euchresta horsfieldii (Lesch) Benn.) is an endangered medicinal plant known for its function as aphrodisiac, antidote, expectorant, and tonic. It also can be used to cure TBC. Research has found that pranajiwa is difficult to be propagated, but the tissue culture propagation method has never been applied to this species. This research is aimed to figure out the procedure to induce callogenesis of pranajiwa with the addition of Benzyl Amino Purine (BAP) and 2,4-Dichlorophenoxyacetic acid (2,4-D) and to determine the proper combination of BAP and 2,4-D to produce an optimal callus. MS-based medium with 25 combinations of BAP and 2,4-D has been used for the callus induction. Parameters observed in this research are callus appearing time, percentage of inducted calluses, and morphology observation, including the texture and color of the callus. Result shows that 90.67% calluses was successfully inducted. The optimal callus was obtained by combining BAP 1 ppm and 2,4-D 1,5 ppm with 5 days of appearing time, friable texture and green color.

​​Effect of Different Media on in vitro Shoot Regeneration from Various Explants of Lilium longiflorum Cv. Elite, Brunello, Cordelia

Background: Lilium (Lilium longiflorum Thunb.) belongs to the family Liliaceae and is a native of Northern Hemisphere (up to South Canada and Siberia). Conventionally, lilium can easily be propagated by sexual and asexual methods of propagation but these prevalent methods are not capable of meeting the increasing demand in domestic and global market. Generally, lilium is propagated through bulbs which has bottlenecks like high cost of planting material, infected from seed borne and other diseases like mosaic, grey mould, etc. So, in vitro multiplication of lilium is utmost important. Lily can be easily regenerated from explants The objective of present study was to identify the best media and the explant among scales and nodal segments belonging to the three different cultivars viz., Elite, Brunello, Cordelia where maximum number of shoots could be regenerated. Methods: The present investigation was carried out in the tissue Culture Laboratory of the Centre for Research and Application in Plant Tissue Culture. Twenty-five media combinations were used to study their effect on in vitro shoot regeneration from various explants of Lilium longiflorum cultivars. Two type of explants viz. scales and nodal segments were aseptically excised from Elite, Brunello, Cordelia cultivars of Lilum and cultured on Murashige and Skoog (MS) medium containing different concentrations and combinations of 6-benzyl amino purine (BAP), kinetin and Naphthalene Acetic Acid (NAA). Result: In cultivars Elite and Brunello, the maximum number of shoots were observed when nodal segments were used on media combination i.e., LM-8 (MS+BAP 3.0 mg/l+NAA0.1 mg/l). In cultivar Cordelia, scales showed the maximum response on media LM-24 (MS+Kinetin 2.0 mg/l+NAA 0.1 mg/l).

Regeneration of Medicinal Plant Solanum virginianum L. Via Somatic Embryogenesis from Seed Explant

Solanum virginianum L., belonging to the family Solanaceae is an important medicinal plant, used in traditional ayurvedic medicines. The present study was aimed to develop an efficient procedure for inducing somatic embryogenesis followed by regeneration in S. virginianum. The seeds of S. virginianum were used as explant and inoculated on ½ MS (Murashige and Skoog) basal medium. Stem explant from in vitro germinated seeds was used for callus induction. MS medium supplemented with 1 mg L-1 BAP (6- Benzylaminopurine) was used for callus induction. Callus proliferation started when initiated callus was sub-cultured on various concentrations and combinations of BAP, IAA (Indole-3-Acetic Acid), IBA (Indole-3-Butyric Acid), 2,4-D (2,4-Dichlorophenoxyacetic acid) and NAA (Naphthalene acetic acid). The highest embryogenesis took place in BAP (@ 2 mg L-1) + IAA (@ 5 mg L-1) combination. The somatic embryos were transferred onto MS basal medium for maturation. These embryos matured and were converted into complete plantlets with well-developed shoot and root system. Highest survival percentage (98±1.24%) was observed on red soil : sand (1:1) potting mixture used for acclimatization.

Effect of plant growth regulators on in vitro induction and maintenance of callus from leaf and root explants of Atropa acuminata Royle ex Lindl

Atropa acuminata, an important medicinal plant belonging to family Solanaceae is under tremendous threat of extinction in its natural habitat due to the overexploitation by pharmaceutical industries. Present study is an attempt of establishing callus cultures of this important medicinal plant as callus has considerable potential as an alternative for production of secondary metabolites for industrial use, hence reducing pressure on natural populations.Callus cultures were established from leaf and root explants of Atropa acuminate. Murashige and Skoog (MS) media containing different concentration and combinations of 6-Benzyl Amino Purine (BAP), Naphthalene Acetic Acid (NAA), Kinetin (Kn) and 2,4- Dichloropheoxyacetic acid (2,4- D) were used for callus induction. Different phytohormonal combinations resulted in different types and degrees of callus. The combination of BAP and NAA on MS media supplemented with 0.5 mg/l BAP in combination with 1.0 mg/l NAA, was found to be the most efficient for in vitro callus development from root explants and from leaf explants most effective combination and concentration was 1 mg/l of both BAP and NAA. The maximum mean fresh weight of callus formed using root explants was 33.13 mg per explant and maximum fresh weight obtained from leaf explants was 22.14 mg per explants.

An Efficient in vitro Propagation Protocol of Pot Calla lily (Zantedeschia spp cv. Orania and Sunclub) via Tuber Production

Zantedeschia spp is an important flower in the ornamental plants market. Due to the high demand for this plant in the horticultural industry, it is indispensable to introduce an in vitro protocol for its mass propagation. For this aim, the tubers of calla lily were disinfected in a hot water bath with different temperatures (30, 35, 40, 45, and 50 oC) and duration (30 or 35 min). Then explants were cultured on MS medium with different combinations of 6-Benzylaminopurine (BAP) and Kinetin (Kin). Based on the obtained results, the highest disinfection percentage (more than 90%) was obtained at 45 oC for 35 min. Also, the highest proliferation rate (with an average of 15.33 and 14.32 in cv. Orania and cv. Sunclub, respectively) was observed in 2.5 mg L-1 BAP + 1.5 mg L-1 Kin. The highest rooting percentage (100% in both cultivars) and root number per explant (with an average of 4.00 and 3.03 in cv. Orania and Sunclub, respectively) was obtained in 0.5 mg L-1 Indole-3-acetic acid (IAA) + 0.1 mg L-1 Kin, but the highest root length (with an average of 120.0 and 106.6 mm in cv. Orania and Sunclub, respectively) was observed in 1.0 mg L-1 IAA + 0.1 mg L-1 Kin. In MS medium + 2.0 mg L-1 Indole-3-butyric acid (IBA) + 4% Sucrose, the highest number of tubers (with an average of 6.66 and 5.21 in cv. Orania and Sunclub, respectively) was formed. The highest fresh and dry weights (with an average of 948.33 and 851.33 mg in cv. Orania and Sunclub, respectively) of tuber were obtained in 2.0 mg L-1 IBA + 6% sucrose. The rooted and tuberous plants were adapted in the greenhouse successfully.

A robust in-vitro protocol for shoot multiplication of Echinacea angustifolia

Echinacea angustifolia is an important medicinal and ornamental plant found in various regions of the world. This species may become threatened due to over-exploitation for medicinal uses. The present study was carried out to develop a rapid in vitro multiplication protocol using explants raised from seedlings. Complete plantlets have been successfully regenerated from shoot tip, leaf, petiole, stem, and root explants. The explants were incubated on Murashige and Skoog (MS) culture medium supplemented with different combinations of 6-benzyl amino purine (BAP) naphthalene acetic acid (NAA). Multiplication of shoot tips was achieved on BAP while from other explants; multiple shoots were regenerated on MS medium supplemented with BAP and NAA. The response of growth regulators was investigated. The shoot tip explants showed the highest rate of shoot induction (80%) and a shoot regenerated about 25 shoots per explants when cultured on MS media supplemented with BAP (4 µM) within 6–7 weeks. The leaf explants also developed shoots and each shoot generated 15 shoots in MS medium supplemented with BAP (5 µM) and NAA (5 µM). Shoot regeneration was also achieved with other explants mainly root, petiole, and stem on MS medium containing BAP (5 µM) + NAA (5 µM). The multiple elongated shoots were obtained on MS basal medium. Well-developed roots were achieved on MS medium in combination with BAP and NAA and 80% of plantlets were survived in the soil successfully in the hardening process. A robust and optimized protocol will be helpful for the mass production of this medicinal plant.

ANALISIS SESAR-SESAR MAJOR SEMENANJUNG MALAYSIA DENGAN TEKNIK GRAVITI DEKONVOLUSI EULER 3D

The application of biotechnology in upland rice improvement programs depends on the availability of efficient regeneration protocols.  Although protocols for shoot regeneration of upland rice are available, none has been reported for pigmented cultivars.  This study reports on a protocol for callus induction and regeneration of Tadong, a pigmented upland rice cultivar from Sabah.  For callus induction, immature embryos were cultured on media containing 2,4-Dichlorophenoxyacetic (2,4-D) at various concentrations (0 – 2.5 mg/L) and on different types of media (MS; MSB5; N6B5; N6).  To induce shoot regeneration, callus explants were cultured on MS medium supplemented with combinations of 6-Benzylaminopurine (BAP) at various concentrations (0 – 3.0 mg/L) and 1-Naphthaleneacetic acid (NAA) at 1.0 mg/L.  To induce shoot development, callus explants were pre-treated with Thidiazuron (TDZ) at various concentrations (0-1.0 mg/l) and exposed to different desiccation periods (0 – 72 hours).  2,4-Dichlorophenoxyacetic at 2.5 mg/L and N6B5 medium resulted in the highest percentages of explant forming callus which were 60.3 ± 17.0 % and 58.7 ± 9.8 % respectively.   The regeneration media failed to induce shoot on callus explants, instead, green spots were formed on the surface of the callus.  The green spots were stimulated to develop into shoots when the callus explants were pre-treated with 0.5 mg/L TDZ or exposed to partial desiccation for 24 h, the percentages of explant forming shoot were 35.7 ± 4.8 % and 47.7 ± 6.8 % respectively.   Shoots developed into complete plants on hormone-free MS medium and acclimatized. 

CALLUS FORMATION AND PLANT REGENERATION FROM IMMATURE EMBRYOS OF PIGMENTED UPLAND RICE (Oryza sativa cv. Tadong)

  The application of biotechnology in upland rice improvement programs depends on the availability of efficient regeneration protocols.  Although protocols for shoot regeneration of upland rice are available, none has been reported for pigmented cultivars.  This study reports on a protocol for callus induction and regeneration of Tadong, a pigmented upland rice cultivar from Sabah.  For callus induction, immature embryos were cultured on media containing 2,4-Dichlorophenoxyacetic (2,4-D) at various concentrations (0 – 2.5 mg/L) and on different types of media (MS; MSB5; N6B5; N6).  To induce shoot regeneration, callus explants were cultured on MS medium supplemented with combinations of 6-Benzylaminopurine (BAP) at various concentrations (0 – 3.0 mg/L) and 1-Naphthaleneacetic acid (NAA) at 1.0 mg/L.  To induce shoot development, callus explants were pre-treated with Thidiazuron (TDZ) at various concentrations (0-1.0 mg/l) and exposed to different desiccation periods (0 – 72 hours).  2,4-Dichlorophenoxyacetic at 2.5 mg/L and N6B5 medium resulted in the highest percentages of explant forming callus which were 60.3 ± 17.0 % and 58.7 ± 9.8 % respectively.   The regeneration media failed to induce shoot on callus explants, instead, green spots were formed on the surface of the callus.  The green spots were stimulated to develop into shoots when the callus explants were pre-treated with 0.5 mg/L TDZ or exposed to partial desiccation for 24 h, the percentages of explant forming shoot were 35.7 ± 4.8 % and 47.7 ± 6.8 % respectively.   Shoots developed into complete plants on hormone-free MS medium and acclimatized.

Chitosan enhances growth and survival rate of in vitro-cultured plantlets of banana cultivar “Grand Naine”

ABSTRACT Banana (Musa spp.) is one of the most popular fruits consumed worldwide. Multiplication of banana through tissue culture is faced with the problem of fungal contaminations. Shrimp chitosan reportedly enhances growth in tissue-cultured plants and reduces fungal and bacterial infections during the in-vitro culture of plants. The present investigation was conducted to establish an in-vitro propagation protocol with zero fungal and bacterial contamination of cultivar ‘Grand Naine’ using explants from the rhizome and sucker. Different combinations of 6-benzylamino purine and indole-3-acetic acid were tested in Murashige and Skoog medium to validate the influence of phytohormones on the response of explants and regeneration of shoots and roots in it. The best response and regeneration were observed at BAP 5.0 mg/L and IAA 0.5 mg/L for the rhizome explants and BAP 4.0 mg/L and IAA 0.5 mg/L for the sucker explants. The best root formation was observed at IBA 0.8 mg/L and 1.0 mg/L for the rhizome and sucker explants, respectively. To validate the effect of shrimp chitosan on the growth of both explants, culture media was supplemented with chitosan in different concentrations. The best growth of banana explants was observed with 25 mg/L shrimp chitosan. The plantlets were hardened successfully in the greenhouse, with a survival rate of 93% for the sucker and 91% for the rhizome explants; compared with 68% and 65% for both positive controls, respectively. The present standardized protocol should be useful for the mass production of banana, germplasm conservation and development of economically important banana cultivars. Abbreviations: BAP: 6-Benzylamino purine; CS: Chitosan; DPI: Days post inoculation; IAA: Indole-3-acetic acid; IBA: Indole-3-butyric acid; MS: Murashige and Skoog medium; MT: Metric ton; Ha: Hectare; mg/L: Milligram per liter

OPTIMIZATION OF SHOOT INITIATION USING RESPONSE SURFACE METHODOLOGY AND CALLUS MEDIA STANDARDIZATION FOR MICROPROPAGATION OF CENTELLA ASIATICA

Centella asiatica, a medicinal herb consumed as a food supplement and used in traditional and folklore medicine is being overexploited for its therapeutic properties. Lack of adequate cultivation and replacement practices could engender the depletion of the wild variety. Application of plant tissue culture has helped in the regeneration and preservation of several plants in-vitro. The present study reports media standardization for micropropagation and callus induction for the conservation of Centella asiatica using field-grown plants. In media standardization for callus induction, appreciable results were obtained where full strength MS (Murashige and Skoog) media was supplemented with 2 mg.L-1 IAA (Indole acetic acid) and a combination of 0.5 mg.L-1 Kn (Kinetin) with 1 mg.L-1 IAA while MS media supplemented with a combination of 0.5 mg.L-1 Kn and 1.5 mg.L-1 IAA was used for callus proliferation. Combination of BAP (6- Benzyl amino purine) and IAA from preliminary experimentation on micropropagation showed high shooting frequency and were further optimized with parameters such as sucrose and incubation period using Response surface methodology (RSM) to study their individual and interactive effects. A maximum shoot height of 3.8 cm was obtained from the optimized values after 13 days with both BAP and IAA at 0.5 mg.L-1 and sucrose at 6% with the R2 value at 0.73.

The Effect of Plant Growth Regulators (PGRs) on Efficient Regeneration of 12 Recalcitrant Indica Rice (Oryza Sativa L.) Genotypes

Biotechnological approaches provide an efficient and expeditious system for developing agronomic features of rice at the cellular and molecular level compared to traditional breeding methods. The in-vitro regeneration method optimization of recalcitrant indica rice genotypes is a critical step for the biotechnological approaches for further development as the optimum concentrations of Plant Growth Regulators (PGRs) for in-vitro regeneration vary from genotype to genotype. Here, 12 important rice genotypes, including local cultivars and varieties, were collected from different regions of Bangladesh and evaluated for their in-vitro regeneration potentiality. Mature dehusked sterilized rice seeds were inoculated on different basal media (MS, N6, B5) supplemented with different concentrations of 2,4-Dichlorophenoxyaceitic acid (2,4-D) for callus induction. Induced calli were transferred to N6 media supplemented with different combinations of 6-Benzylaminopurine (BA) and 1-Napthaleneacetic Acid (NAA) for shoot induction. Regenerated shoots were rooted on half-strength MS media without supplementation of any hormones. Treatments of different concentrations and combinations of plant growth regulators showed varying results. Maximum callus induction frequencies were observed with 2.5-3.0 mg/l supplementation of 2,4-D on N6 media where Saat, Jamaibabu and Nimikhanai showed the highest callus induction frequencies (84.44%; 78.89 and 80% respectively) and maximum shoot initiation was observed at 2.5 mg/l BA +1.0 mg/l NAA and 3.0 mg/l 6-BA +1.5 mg/l NAA supplemented media where rice genotypes BRRI Dhan71, Nayapaijam, Nimikhanai and BRRI Dhan28 showed the highest shoot induction frequencies-86.67, 85.00, 86.67 and 88.33% respectively. Nearly all of the induced shoots showed root formation on half-strength MS media. This study demonstrates variations in regeneration response of the rice genotypes where cultivars Jamaibabu, Nayapaijam and Nimikhanai showed the highest in-vitro regeneration potential.

In vitro spore germination and early gametophyte development of Cibotium barometz (L.) J. Sm. in different media

Abstract. Isnaini Y, Praptosuwiryo TNg. 2020. In vitro spore germination and early gametophyte development of Cibotium barometz (L.) J. Sm. in different media. Biodiversitas 21: 5373-5381. Cibotium barometz (L.) J. Sm. is known as the golden chicken fern and included in Appendix II of CITES. It is an important export commodity for traditional and modern medicine. Globally, populations of this species are under significant pressure due to overexploitation in the wild. In vitro culture is one of the technologies used for ex-situ propagation and conservation of rare and endangered ferns and lycophytes. This study’s objectives were: (i) to observe in vitro spore germination and early gametophyte development of C. barometz, and (ii) to determine the best culture medium for rapid spore germination and early development of the gametophytes. The sterilized spores were sown in half-strength Murashige & Skoog (½MS) basal medium supplemented with combinations of 6-Benzylaminopurine (BAP) and α-Naphthalene acetic acid (NAA). A factorial combination of four BAP concentrations (0, 2, 4, and 6 mg L-1) with four concentrations of NAA (0; 0.01; 0.03 and 0.05 mg L-1) created 16 treatments replicated in a Completely Randomized Design. Spore germination of C. barometz was observed to be Vittaria-type, and its prothallial development was Drynaria-type. Spore germination started 7-14 days after sowing. Young heart-shape gametophytes consisting of 110-240 cells were formed in 45-61 days after sowing. The two best spore culture media for rapid spore germination and development of C. barometz gametophytes were ½ MS with or without 2 mg L-1 BAP.

English

Vegetatively propagated crops are exposed to pests every growing season, providing them with the opportunity to feed on the plants and transmit diseases. Meristem culture is an important method to clean for viruses to restore high yields. Media optimizations with different concentrations and combinations of 6-benzylamino purine (BAP) and Naphthalene Acetic Acid (NAA) were tested for their effects on shoot initiation from meristems. This was followed by regeneration and shoot multiplication from nodal cuttings in five elite varieties of sweet potato. There was a highly significant genotype (variety) x environment (media) interaction for all experiments. Concentrations and combinations of BAP and NAA significantly affected the percentage of meristem survival/regeneration and subsequent shoot multiplication. The best combination of the plant growth regulators was NAA (0.1 mg/L) and BAP (1.0 mg/L). For further propagation, shoot numbers per single nodal cuttings were significantly affected by BAP concentrations (p < 0.05) and genotypes. The medium which gave maximum numbers of shoots was ½ MS media supplemented with BAP (2.5 mg/L) and GA3 (0.5 mg/L). The tallest shoots, with the highest number of internodes, suitable for transfer to soil, was obtained from ½ MS media without any growth regulators for all of the cultivars.   Key words: Media, meristem, multiplication, regeneration, shoot, sweet potato.

Effect of benzyl amino purine and 1-methylcyclopropene in maintaining rooting quality of chrysanthemum (Chrysanthemum morifolium Ramat cv. ‘White Fiji’) cuttings

The quality of chrysanthemum cuttings is influenced by ethylene. During postharvest storage, ethylene accelerates the quality loss of cutting by enhancing chlorophyll degradation. Application of 1-Methylcyclopropene (1-MCP) or Benzyl Amino Purine (BAP) can be used as an approach to inhibit unfavourable ethylene effects. The objective of this study was to find out the effect of a combination of 1-MCP or BAP and storage period on the quality of chrysanthemum cuttings. The experiments were arranged using a completely randomized design consisting of nine treatments. The cuttings were treated with 1-MCP 700 nL/L or BAP 2.5 mg/l. After treatment, they were stored for 0, 3 and 7 days, then the cuttings were rooted in soilless culture for four weeks. The results showed that 1-MCP or BAP maintained leaf freshness and inhibited chlorophyll degradation on cuttings at 3 and 7 days of storage period. However, the application of 1-MCP slightly inhibited root growth compared to that of untreated cuttings. Interestingly, the application of BAP effectively reduced the cuttings damage for seven days and did not cause the inhibition of root growth (weight of fresh root, root number, root length and root volume) that was comparable to that of untreated cuttings. These results collectively showed that BAP was more effective than 1-MCP in maintaining rooting quality of chrysanthemum cuttings.

Callus induction in hydnocarpus pentandra (Buch. Ham.) Oken using shoot tips

Hydnocarpus pentandra (Buch. Ham.) Oken is a Vulnerable medicinal tree found in Western Ghats of India. It is highly exploited due to its medicinal properties and it is mainly used in leprosy treatment. An experiment was undertaken at Division of Plant Genetic Resources, ICAR-Indian Institute of Horticultural Sciences, Bengaluru during 2018-19. The experiment was laid down in Completely Randomized Design (CRD) with 16 treatments and three replications. Shoot tips from young seedlings were used as explant and treated in MS media with a different hormonal combination like Benzyl Amino Purine (BAP), Indole Acetic Acid (IAA), Kinetin and Thiaduzerol (TDZ). Among the treatments, highest shoot length (2.06 cm) and maximum number of shoots produced per explant (3.86) were found in the treatment combination of BAP (1.0 mg/l) and TDZ (0.5 mg/l) followed by the treatment combination of BAP (2.4 mg/l) and IAA (0.05 mg/l). Few treatments showed callus formation but no shoots were developed in these treatments.

Optimization of Plant Regeneration in Different Pepper (Capsicum annuum L.) Lines

Development of an efficient plant regeneration protocol is essential for vegetable crop advancement by biotechnological methods. In this study, regeneration protocols of four pepper lines of different pepper types were optimized. Different protocols for organogenesis were investigated in regeneration experiments. Optimum plant regeneration was obtained in different combinations of 6-Benzylaminopurine (BAP) and Indole-3-acetic acid (IAA) in organogenesis assays. In organogenesis experiment, the highest shoot development was determined as 80% from hypocotyl explant of Demre pepper in 4 mg L-1 BA+0.5 mg L-1 IAA, 80% from hypocotyl explant of Charleston pepper in 3 mg L-1 BA+0.5 mg L-1 IAA, 80% from hypocotyl explant of capia pepper in 5 mg L-1 BA+0.5 mg L-1 and 84% from hypocotyl explant of bell pepper in 5 mg L-1 BA+0.5 mg L-1 IAA. Afterward, shoots were rooted and whole plants were obtained.