The Effect of Polyethylene Glycol (Peg) on Calluses of Sweet Basil (Ocimum Basilicum L.) (original) (raw)
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anthers were taken from bean flower buds of the variety Aquadlce and microspore developmental stages were determined according to flower bud sizes. Anthers were cultured on modified Schenk and Hildebrandt medium (SH) supplemented with EDTA ferric monosodium salt to the SH medium which was taken from formulation of Murashige and Skoog (MS), casein hydrolysate 400 mg/l, some amino acids, and different combinations of growth regulators; kinetin and naphthalene acetic acid and 2, 4- dichlorophenoxyacetic acid (Kin + NAA + 2, 4-D) and benzyl adenine (BA + NAA + 2,4-D) in different concentrations. The combination (BA + NAA + 2, 4-D) used with flower bud sizes (7.1-8) mm, containing uninucleate microspores, gave better callus induction (50%). It was also found that the concentrations [1.0, 2.0] mg/l of ascorbic acid were better to prevent the accumulation of phenols in the medium than the concentration [0.0] mg/l. The cytological analysis revealed that the number of chromosomes in callus induced from flower bud sizes (2.5-6.4) mm contained diploid (2n=12 chromosomes), whereas chromosomes in callus induced from flower bud sizes (6.5-8.0) mm contained haploid (n=6 chromosomes) with the presence of some diploid cells (12 chromosomes).
Phytochemistry, 2012
Metabolic changes were studied, which accompanied the conversion of 6 month old HiII maize nonregenerable (NR) calli into regenerable (R) calli when cultured for 63 days with 10% polyethylene glycol (PEG) (3350 MW) in culture medium. The conversion of 6 month old NR to R callus morphotype caused by PEG application decreased cell wall contents in callus dry mass and changed cell wall phenolics making their profile similar to that of R callus by reduction of lignin and ester-and ether-bound phenolic concentrations, including p-coumaric acid and ester-and ether-bound diferulates and by increase of the ratios of ester-and ether-bound ferulic acid/coumaric acid and ferulic acid/diferulic acid in cell walls of NR callus. Some similar changes of cell wall phenolics caused by PEG application were also found in 48 month old NR callus, that changed the morphology, but did not regenerate plants. However, there were no changes in the old callus in levels of total ester and ether-bound cell wall phenolics and substantially smaller decreases were found in ratios of ester-and ether-bound ferulic acid/coumaric acid and ferulic acid/diferulic acid, as well as in diferulate concentrations compared to young NR callus cultured with PEG. Remarkably, application of PEG also changed the primary metabolism of young NR callus tissues, so that they acquired metabolic features of highly regenerable callus. These data clearly suggest that PEG alters metabolism of NR calli, so they acquire biochemical characteristics of R calli, and that adaptive osmotic adjustments vary in different types of callus tissues.
STANDARDIZATION OF FRIABLE CALLUS DEVELOPMENT IN CATHARANTHUS ROSEUS (LINN.) G. DON Original Article
Objective: The objective of the study was to develop an effective hormonal combination for the maximum growth of callus and development of friable calli using the same medium with reduced concentration of agar. Methods: The percentage responses of five varied growth hormonal combinations and concentrations, supplemented with Murashige and Skoog (MS)medium were recorded. The effect of casein hydrolysate on callus induction was also studied. The nature of friable calli obtained from best responsive media fortified with 0.7% and 0.6% agar was observed. Results: The present study revealed that, three media viz., MS + 1.0 mg/L BAP + 1.0 mg/L NAA, MS + 1.5 mg/L 2,4-D + 1.0 mg/L Kin and MS + 1.5 mg/L 2,4-D + 0.5 mg/L BAP, as the best responsive media in the descending order. The effect of casein hydrolysate supplemented along with the above three media revealed MS + 1.0 mg/LBAP + 1.0 mg/L NAA + 1.0 gm/L casein hydrolysate as the best responsive media. Also, the above media supplemented with 0.6% agar was found to be the effective in terms of nature and amount of friable callus obtained. Conclusion: The results indicated MS + 1.0 mg/L BAP + 1.0 mg/L NAA + 1.0 gm/L casein hydrolysate + 0.6% agar (85% response) as the best media for the growth and development of both callus and friable callus.
Indian Journal of Scientific Research, 2020
Tissue culture study was performed to standardize the technique and select suitable plant growth regulators and their concentrations for efficient callus induction from the nodal explants of Asparagus racemosus willd, an important medicinal herb. Different concentrations of NAA, 2,4-D, BAP and KN were supplemented in MS basal medium in which the nodal explants were inoculated. It was observed that MS + 0.2 mg/l NAA was the best culture condition in which maximum percentage of response 90.36 was obtained. This was followed by the culture conditions that were MS+ 2.0 mg/l where the percentage response was 88.68%. Similarly, next higher percentage 88.62 was observed in MS+1.0 mg/l BAP + 1.0 mg/l NAA. It was further observed that growth rate was excellent where the percentage response was higher. Second it was noted that the calli had both compact and friable texture and their colour varied from green yellow to brown even where the growth was excellent. It was further noted that NAA or BAP had no response for callusing when used alone. However, KN at higher concentration 2.0 mg/l gave the highest percentage of response for callusing. NAA + BAP and NAA + KN at the similar concentrations gave different result. In the MS medium having 1.0 mg/l BAP + 1.0 mg/l NAA the percentage of response was 88.62 while MS + 1.0 mg/l KN + 1.0 mg/l NAA the percentage of response was 62.34 only. Growth rate of the calli also differed in different culture conditions.
Agro-Science, 2007
Growth of callus explants was studied by manipulating the hormonal composition of Von Arnold and Eriksson's (AE) 1977 medium. Various auxin-cytokinin combinations were tested. Growth of callus on fresh weight basis, was best promoted by 2mgl-1 Naphthalene Acetic Acid (NAA) and also by the interaction between Indole Butyric Acid (IBA) at 2mgl-1 and 6-Benzyl Amino Purine (BAP) at 0.5mgl-1. Although there were no significant differences among the various combinations of NAA levels with kinetin levels and NAA levels with BAP levels with respect to fresh weight production, per cent increments over the controls were determined. Maximum increments were obtained from 1mgl-1 NAA x 0.5mgl-1 kinetin (406.5%) and 1mgl-1 NAA x 0.5mgl-1 BAP (198.06%). There were strong indications that high fresh weights were not always synonymous with dry matter production. Among the levels of NAA, 0mgl-1 and 0.5mgl-1 produced the highest percentages. In NAA x kinetin cultures, 0mgl-1 produced a value of 0.467% while 0.5mgl-1 in NAA x BAP cultures produced a value of 0.485%. For kinetin levels, 5mgl-1 produced the maximum percentage of 0.451% while for BAP levels, 0mgl-1 (0.471%) and 5mgl-1 (0.462%) produced the optimum percentages. With respect to the hormonal interactions, optimal percentages were obtained in 0mgl-1 NAA x 5mgl-1 kinetin (0.643%) and 0.5mgl-1 NAA x 0mgl-1 BAP (0.625%) cultures. Spectrophotometric studies revealed the existence of an association between the chlorophyll content of calluses and the grade of greening. Callus greening was best promoted by the main effects of kinetin (5mgl-1), BAP (5mgl-1) and NAA (2mgl-1). An unusual sign of physiological specialization (reddish pigmentation) was observed in some of the callus clumps. The implications of the results have been discussed.
2015
Labisia pumila (BI.) F. Vill (Myrsinaceae) also known as Kacip Fatimah is a medicinal plant with an enormous therapeutics value and used widely for herbal industries in Malaysia. This study reports the induction of globular calli from stem explants of L. pumila var. alata. The embryogenic characteristics of the globular calli were further confirmed by the morphological and histological analyses using the light microscope of sectioned material and scanning electron microscope. Globular callus was induced from stem explants that were pre-treated/un-pretreated with thidiazuron (TDZ) on Murashige and Skoog (MS) medium supplemented with 20 µ M 2,4-dichlorophenoxyacetic acid (2,4-D) and various amino acids (glutamine, arginine, proline and leucine) at the concentrations of 0.05, 0.1, 0.25 and 0.5 mM. After six weeks of incubation, explants pre-treated with TDZ induced 100% of globular callus while only friable calli were observed on un-pretreated explants. The addition of amino acids at c...
Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 2013
Callus holds great potential for biotechnology applications in plants. In this study, several experiments were performed to establish a protocol for in vitro induction of good quality callus by using immature cotyledons and embryos of Juglans regia cv. ‘Xiangling’ as the explants. DKW basal media supplemented with different concentrations of plant growth regulators were applied to determine the induction rate, texture and color of the recovered callus. Results showed that the optimum culture medium for the callus induction from these explants was a DKW basal medium consisting of 1 mg/L BA, 2 mg/L KT 250 mg/L glutamine, 500 mg/L casein hydrolysate, 200 mg/L Vc, 50 g/L sucrose and 2.8 g/L Gelrite (i.e., the T8 medium). The average rate of callus induction on the medium reached 80.7% for the immature cotyledons and 66.7% for the immature embryos, and all the callus was characterized by a friable, creamy or yellow appearance. In vitro proliferation of callus was not very successful ...
2019
Devegulu hollyhock is used by public for the treatment of certain diseases. This study investigated the appropriate protocols for in vitro callus formation, regeneration and micropropagation of various explants of 14-day and 28-day old Devegulu seedlings germinated from the seeds in a sterile environment. Different concentrations of 2,4- dichlorophenoxy acetic acid (2,4-D) were used as plant growth regulator and Murashige and Skoog basal medium was used as basic medium. Regenerated plantlets were firstly transferred to rooting medium to promote root formation, after that they were planted into soil. Both 14-day and 28-day explants produced a higher rate of callus in MS medium including 2,4-D auxin hormone. The regenerated plantlets were obtained from the shoot tip and node explants of 28 days old seedlings after incubation in MS medium for 6 weeks. Direct regeneration from the node and shoot tip was observed in this hormone free medium. Micropropagation rates were 44.4 % and 12.5 % ...
IN VITRO INDUCTION OF CALLUS FROM FOLIAR EXPLANTS IN ROCOTO (CAPSICUM PUBESCENS RUIZ & PAV.)
Rocoto (Capsicum pubescens Ruiz & Pav.) is a native plant of Peru, used in the national gastronomy with great nutritional value; it has pharmaceutical and medicinal properties. The crop is susceptible to diseases caused by phytopathogens, which are spread by seeds of infected cultivars or seedlings. Through the use of biotechnological tools such as in vitro plant tissue culture, it is possible to obtain plants free of pathogens, of good quality and great agronomic potential. Therefore, the present work aimed to develop a methodology for the in vitro induction of callus from hot pepper leaves. In vitro germinated rocoto seedlings were used, from which the first true leaves were selected, which were sectioned into 1 cm explants and placed in different MS culture media added with 2,4-dichlorophenoxyacetic acid (2,4-D). The induction response to calllogenesis was evaluated in five treatments with different concentrations of 2,4-D (0, 0.25, 0.5, 0.75 and 1 mg l1), under conditions of 25 C in the dark for 35 days. The culture media added with 0.75 and 1 mg l1 of 2.4-D allowed to obtain 100% induction of calluses in the hot pepper leaves with 81% and 86%, respectively of grade 3 callus formation. This study, pioneer for the species, is good for the potential use of breeding programs.