scholarly journals Establishment of callus induction system, histological evaluation and taxifolin production of Larch

Author(s):  
Xinglong Liu ◽  
Yingchun Zhao ◽  
Xueyan Chen ◽  
Ling Dong ◽  
Yinan Zheng ◽  
...  

Abstract Taxifolin in larch is an important natural active ingredient, which prevents and treats liver diseases and cardiovascular diseases, and has many health benefits and application values ​​such as anti-tumor, anti-virus, and anti-aging. However, the traditional extraction method is not conducive to the protection of the ecological environment, and also limits the industrial production of taxifolin from larch. Therefore, the purpose of this research is to establish a larch callus culture system and obtain taxifolin from callus. The shoots of the stem-tips of Larix olgensis were used as explants to be cultured. The light conditions, optimal medium and hormone ratio for callus induction were screened to evaluate the callus induction characteristics of larch and its influence on the accumulation of taxifolin. The results showed that: callus formation started on 28–51 days, and the best conditions for inducing callus were explored: 12 h/d light culture MS + 2,4-D 2 mg/L + 6-BA 1 mg/ L + KT 0.1 mg/L; the callus induction rate is 50%-80%; the highest condition for producing taxifolin 6-BA 1 mg/L + NAA 0.1 mg/L, reaching 0.414%. In conclusion, this study represents a suitable induction condition for larch callus culture and taxifolin accumulation.

Food Research ◽  
2020 ◽  
Vol 4 (S5) ◽  
pp. 115-123
Author(s):  
A.N. Salihan ◽  
N.A. Yusuf

Ananas comosus var. MD2 is currently the most preferred pineapple variety in the international market due to its pleasant aroma and high Brix acidity ratio. In vitro approaches such as callus culture is promising in producing disease-free plantlet. However, there are limited studies reported on callus culture of MD2 variety despite the potential of in vitro regeneration through biotechnological advances. The purpose of the study was to determine the effect of plant growth regulators (PGRs) i.e., 2,4- dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BAP) and Thidiazuron (TDZ) on callus induction from leaf explant of MD2 pineapple. Leaf base explants were cultured on Murashige and Skoog (MS) media supplemented with varying concentration of 2,4-D (0.5 to 6.0 mg/L) alone and in combination with BAP (1.0 to 3.0 mg/L). The frequency of callus induction was seen significantly highest (91.67±8.33%) with maximum callus fresh weight (0.25±0.07 g) at a combination of 2.0 mg/L 2,4-D and 2.0 mg/L BAP. The shortest duration of callus formation was seen on day 12 with the lowest concentration of 2,4-D, 0.5 mg/L. There is a moderate correlation between the earliness of callus formation and the frequency of callus induction (P<0.01). The most favourable media for callus proliferation was 6.0 mg/L 2,4-D and 2.0 mg/L TDZ as the highest fresh weight of 1.52±0.03 g was recorded. Callus culture has the potential to be a source of plant material and secondary metabolites production. In this study, 2,4-D and BAP have successfully induced callus in MD2 pineapple.


2020 ◽  
Author(s):  
Chongwen Dai ◽  
Yang-yang YAN ◽  
Yumin Liu ◽  
Ya-min LIU ◽  
Yuan-wei DENG ◽  
...  

Abstract Background Landscape industry has been recognized as the "eternal sunrise industry", however, the introduction and cultivation of colored-leaf trees in southern China is relatively vacant.Results The results showed that the induction effect of the stem was the best for the embryogenic callus, and the regenerated plant with genetic stability was obtained. The MS (Murashige&Skoog) +0.8 mg/L TDZ (Thidiazuron) +1.0 mg/L 6-BA (6-Benzylaminopurine) +0.5 mg/L NAA (1-naphthlcetic acid) +35 g/L sucrose+7.5 g/L semi-fixed medium was the best for callus formation. When selecting type VI callus as embryonic callus induction material, MS+0.6 mg/L TDZ+0.5 mg/L 6-BA +2.0 mg/L IAA (Indole-3-acetic acid) +35 g/L sucrose+7.5 g/L semi-fixed medium can get embryonic callus. The optimal medium for adventitious bud induction is MS+1.0 mg/L TDZ+3.0 mg/L 6-BA+0.2 mg/L NAA+1.2 mg/L IAA+35 g/L sucrose+7.5 g/L semi-fixed medium.The induction rate of adventitious roots in MS+0.6 mg/L TDZ+1.0 mg/L 6-BA+3 mg/L NAA+35 g/L sucrose+7.5 g/L semi-fixed medium was the highest, reaching 76%.Conclusion In the course of our research, we found that PGRs play an important role in the callus induction stage, and the effect of TDZ is particularly obvious; The callus cells grow and proliferate according to the "S" growth curve, and can be subcultured when the highest growth point is reached to maintain the rapid proliferation of the callus cells and to avoid inactivation of callus caused by tight niche.


Plants ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 1436
Author(s):  
Xianbao Deng ◽  
Yaqian Xiong ◽  
Jing Li ◽  
Dong Yang ◽  
Juan Liu ◽  
...  

The lotus (Nelumbo nucifera) is one of the most popular aquatic plants in Asia, and has emerged as a novel model for studying flower and rhizome development, and primary and secondary metabolite accumulation. Here, we developed a highly efficient callus induction system for the lotus by optimizing a series of key factors that affect callus formation. The highest efficient callus production was induced on immature cotyledon and embryo explants grown on Murashige and Skoog (MS) basal medium containing an optimized combination of 3 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg/L 6-benzylaminopurine (6-BA). In addition, lotus callus induction was proven to be influenced by lotus genotypes, light conditions, the developmental stages of explants and the time of explant sampling. Collecting immature cotyledons from seeds of the genotype “Shilihe 1”, at 9 days post pollination, and to culture the explants in darkness, are proposed as the optimum conditions for lotus callus induction. Interestingly, highly efficient callus induction was also observed in explants of immature embryo derived aseptic seedlings; and a small amount of lotus benzylisoquinoline alkaloid (BIA) and obvious expression of BIA biosynthetic genes were detected in lotus callus.


2016 ◽  
Vol 18 (1) ◽  
pp. 142-148 ◽  
Author(s):  
M.R.A. SANTOS ◽  
M.C.M. GUIMARÃES ◽  
E.S. PAZ ◽  
G.M.O. MAGALHÃES ◽  
C.A. SOUZA ◽  
...  

ABSTRACT Piper permucronatum is a perennial shrub, a medicinal plant native to the Amazon Rainforest. Traditionally, the tea of its leaves is used to combat menstrual and intestinal cramps, stomach pain, digestive problems, diarrhea, hemorrhage, and nausea. Its leaf’s essential oil is effective against Aedes aegypti larvae; its flavones and flavanones have a fungicidal effect against Clamidosporium cladosporioides and C. sphaerospermum; its hexanic extract is effective against Leishmania amazonensis. The objective of this study was to provide a protocol for callus induction from P. permucronatum leaves and an identification of the callus growth pattern, focusing on the deceleration phase, when the callus cells must be subcultured into liquid medium in order to produce a cell suspension cultures. Leaf explants were inoculated in a solid MS medium supplemented with factorial combinations of 2,4-D, BA, NAA and GA3. Callus formation was evaluated weekly until the 49th day. Subsequently, new explants were inoculated at the hormonal combination that resulted in the highest callus cell proliferation and, every seven days during a period of 70 days, samples were dried and weighed to determine the callus growth pattern. NAA and GA3 were not effective for callus induction. Combinations of 2,4-D and BA resulted in callus induction and proliferation. The highest percentage of callus induction was observed with the combination of 4.52 µM 2,4-D and 4.44 µM BA. The calluses thereby produced were friable and whitish. The callus growth pattern followed a sigmoid shape. The deceleration phase started on the 56th day of culture.


2012 ◽  
Vol 12 (2) ◽  
pp. 93 ◽  
Author(s):  
Budi Winarto ◽  
Nurhayati Ansori Mattjik ◽  
Agus Purwito ◽  
Budi Marwoto

Improvement of selected induction culture media on callus induction in anther culture of anthurium and a histologicalstudy on its callus formation were studied at the tissue culture laboratory of the Indonesian Ornamental CropsResearch Institute from February to October 2008. The objectives of the study were to optimize selected media forcallus formation, reveal cell origin of callus derived from anther culture and shoot formation process. Selectedmedia improved in the study were 1) MMS-TBN containing 0,5 mg/l TDZ, 1,0 mg/l BAP and 0,01 mg/l NAA (Winartomedium, WM) and 2) MMS III supplemented with 1,5 mg/l TDZ, 0,75 mg/l BAP and 0,02 mg/l NAA (Winarto andRachmawati medium, WRM). Improvement treatments were carried out by omission and application of 2,4-D in 0.5mg/l and reduction of medium strength of full, half, quarter, one eighth, one sixteenth, and zero strength. Afactorial experiment was arranged using a randomized complete block design with four replications. Results ofthis study indicated that the highest callus induction was clearly established in WRM. The medium stimulatedpotential growth of anther (PGA) up to 81% with 49% of percentage of anther regeneration (PAR) and 2.7 number ofcallus formed per replication (NCF). Significant improvement in callus formation was also recorded by reduction ofmedium strength of WRM to one eighth compared to others. The reduction induced PGA up to 58% with 29% of PARand 1.8 NCF. From histological studies it was well recognized that regenerated callus on half anthers cultured wasoriginated from middle layer cells of anther wall. The morphogenic response of anther wall cells caused primarilyon no androgenesis effect in microspore cells.


2021 ◽  
Vol 21 (2) ◽  
pp. 427
Author(s):  
Fajri Marisa ◽  
Lisna Hidayati ◽  
Aries Bagus Sasongko ◽  
Tri Rini Nuringtyas

Gyrinops versteegii is an endemic plant in eastern Indonesia that produced agarwood with high quality and economic value. This plant has been threatened by overexploitation which leads on decreasing in the natural population. This research aimed to induce in vitro callus formation to support the sustainable utilization of G. versteegii, which in the end, may support plant productivity. Callus induction was investigated using cotyledon and cultured on MS medium supplemented with several combination of plant growth regulators. The observations were done for the number of explants turned into callus and the duration for the first callus formation. The results showed that combination of 3 mg/L NAA + 0.5 mg/L BAP was recorded as the best combination for callus induction (63.63%). Callus with friable structure and bright color are obtained within nine days of incubation and showed the characteristic of embryonic callus. This result is expected to give a significant opportunity to conserve the natural population of G. versteegii.


Aquilaria malaccensis Lam. and Aquilaria subintegra Ding Hou belong to the family of Thymelaeaceae which is commonly known as gaharu or agarwood. It is a commercially important tree and identified as a potential aromatic plant. The overwhelming responses in the lodging sector reduce gaharu species in the forest. Mass propagation through plant tissue culture technology will substitute this problem. The present study was conducted to investigate the embryogenic callus induction between these two species. The most optimum sterilization method for both species was sodium hypochlorite 5.0% which gave the highest percentage of aseptic culture (95%) with the absence of tissue browning. The leaves of both species were cultured on Murashige and Skoog, (1962) (MS) media supplemented with combination of various concentrations of 6-benzylaminopurine (BAP) (0.5, 1.0, 2.0 and 2.5 mg/L) and 2,4-dichlorophenoxyacetic acid (2, 4-D) (0.5, 1.0, 1.5 and 2.0 mg/L) and kept under dark condition. The explants produced embryogenic, white and compact callus at the end cut of the explants after two weeks of culture in all treatments. The highest frequency of embryogenic callus formation was observed in explants cultured on 2.0 mg/L BAP and 0.5 mg/L 2,4-D for both species. From the present study, the optimum sterilization technique and embryogenic callus induction for A. malaccensis Lam. and A. subintegra were established.


Author(s):  
Aananthi. N

Five rice cultivars viz., ASD 16, White Ponni, Pusa Basmati 1, Pusa Sugandh 4 and Pusa Sugandh 5 belonging to subspecies indica were compared for its ability in callus formation and regeneration. In this experiment, the different parameters viz., the effect of hormones (2,4-D and kinetin), organic supplement (coconut milk O1-CM 100 mll-1, O2-CM 75 mll-1, O3-CM 50 mll-1), explants (seed and immature embryo), media (MS and N6), carbon source (sucrose and maltose) using five genotypes on callus response was studied. The effect of hardening methods was also assessed. Results showed that for enhanced callus induction was with MS medium supplemented with 2.0 mgl-1 2, 4-D + 0.5 mgl-1 kinetin + 30 gl-1 maltose irrespective of explants used. Addition of 100 ml l-1 coconut milk was found have improvement in callus response. The performance of immature embryo was better than seed for callus induction, emrbyogenic callus formation, rhizogenic callus formation and regeneration. MS media provided superiority over N6. Among the genotypes Pusa Basmati 1 rendered outstanding performance in callus behavior. The treatment combination MS + 2.5 mgl-1 BAP + 0.5 mgl-1 NAA + 1.0 mgl-1 KN gave the highest organogenesis response and regeneration of plantlets. Hardening in mist chamber was recognized as the best method to give the highest per cent of regenerated plant lets.


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