The role of carbon sources in phenolic secretion during Cotton Culture is not well understood. The medium can cause the production of phenolic compounds to adversely impact the cotton’s callus culture and ability to regenerate. To overcome this problem, the current study examined the effects of carbon sources on phenolic production in SVPR-2 cotton callus culture.
This study provides a method for controlling Cotton Culture
Be evaluated for high frequencyCallus culture using reduced phenolic production in cotton (Gossypium HarsutumL.) cv. SVPR-2. A protocol was devised to reduce excessive phenolic compounds secretion in cotton callus culture. Callus induction and proliferation are optimized to achieve high frequency callus cultures due to cotton’s naturally high levels of phenolic compounds. To control the phenolic secretion of callus cultures, different carbon sources, such as sucrose, glucose, sucrose, fructose and maltose, were tested at various levels. The best carbon source to control phenolic secretion was 3% maltose. MSB5 medium, which was supplemented with 2,4-D (0.90mM), and Kinetin (4.60mM) both from cotyledon explants, had a high frequency of callus formation. Callus induction with hypocotyl explant was 94.90%, followed by cotyledon (85.20%) MSB5 medium with 2,4-D (0.45 mgmM) and 2iP (2.95 mgmM) produced a tremendous growth of callus with a high response rate. Different hormone treatments produced different colors and textures.
Phenolic production in cotton callus culture. It does this by adjusting carbon sources and evaluating the use of plant growth regulators to improve callus culture efficiency of SVPR-2. China is the largest producer followed by India and the US. Gossypium (hirsutum) is grown in over 90% of all agricultural fields for commercial production. Many biotic and abiotic factors can greatly influence the quality and yield of cotton fibers. These factors can have a huge impact on the yield and growth of the plant, which can lead to economic losses. The most widely-used method for creating transgenic cotton plants that can thrive in adverse environments is genetic engineering. These problems have been solved by many methods. In vitro regeneration is the most effective method to create transgenic cotton plants. Incompatibility barriers make genetic improvement of cotton difficult and takes longer to create desired characteristics. This issue can be avoided by in vitro cultivation of cotton.
It is possible to obtain desired traits for cotton plants within a specified time. In vitro manipulation of cotton is difficult because it is often resistant to in vitro manipulation [28]. There have been many attempts to improve the efficiency of callus culture and regenerate cotton plants for various cotton cultivars around the globe. Many factors, including carbon sources, plant growth regulators and explants, as well as culture conditions, culture vessels, and phenolic compound release, can have a positive or negative impact on the cotton callus cultivation. The main problem that adversely affects the in vitro cultivation of cotton is phenolic compound production [8], [14]. To avoid excess phenolic compound secretion in the culture medium, it is necessary to add a suitable carbon source. A protocol for efficient regeneration is also required to obtain transgenic lines from local cotton varieties. Cotton regeneration is highly dependent on the genotype. In vitro regeneration of cotton plants from local cultivars has been possible. It is therefore important to develop a reliable regeneration protocol in order to produce superior cotton genotypes.
- hirsutum seeds cv. SVPR-2, were obtained at the Cotton Research Station, Tamil Nadu Agricultural University in Srivilliputtur Tamil Nadu, India.
The seeds were soaked in concentrated sulfuric acid, and then washed for 15 minutes under running water. Next, the seeds were washed with Teepol and left to soak for 15 minutes. After three washes with sterile water, the seeds were dried. After 50 seconds of 70% ethanol, the seeds were disinfected for five minutes with 0.1% mercuric chlorineide. Intermittent washes in sterile water were then performed. The seeds were then germinated aseptically using half-strength MS Basal Medium [19]. Inoculated seeds were kept in darkness at 25 +-2 degC for two days before being moved to 16:8h light/dark with light provided by Philips, India cool white fluorescent lamps at 80 mmol m-2/s-1.
Influence of carbon sources Cotton Culture
To evaluate the effects of carbon sources like fructose and glucose on callus formation efficiency and phenolic compound production, we tested them at various concentrations (between 10 and 50 g/l). MSB5 medium was used to culture hypocotyl and cotyledon explants. After two weeks of cultivation, the results were confirmed. The best carbon source was chosen for further research.
Callus induction
Callus induction was done using hypocotyl and cotyledon explants. In vitro-raised seedlings, explants were removed at seven days of age. The cotyledon explants had to be cut into small pieces and the hypocotyls were then cut into 5mm lengths. They were kept on MS salts with B5 vitamins and MS salts fortified with different concentrations of auxins or cytokinins. For callus induction, the explants were placed into a petri plate measuring 15 x 90mm. It contained approximately 30 ml of medium. MS medium supplemented with different concentrations of auxins such as 2,4-D (2,4-Dichlorophenoxyacetic acid), NAA (Naphthalene acetic acid), IBA (Indole-3-butyric acid) and IAA (Indole-3 acetic acid) was initially tested for callus induction. After 2 weeks of culture, the results were visible. The highest concentration of auxin was used as a standard. It was then tested with other cytokines such as BAP (6 Benzyl ampurine), 2iP (2 -Isopentenyl adenine) and KIN (10 Kinetin). For callus formation, cultures were incubated at 25 +-2 degC under cool fluorescent lighting with a 16/8 h light/dark cycle.
Callus proliferation Cotton Culture
For callus proliferation, well-developed calli were removed and transferred to MSB5 medium, which was supplemented with auxins (2.4-D, NAA, and IAA), and cytokinins (2iP. TDZ, and KIN). After 2 weeks of culture, the results were visible.
The study was based on a completely random design that included ten replications. Each experiment was repeated three times. Each petridish contained 10 plants, and there were 100 total explants in each treatment. At every stage Cotton Culture of the analysis, control was maintained. One-way analysis (SPSS version 17) was used to determine the statistical significance of data from this study. Duncan’s multiple range test (p 0.05), and the percentage response were scored using DMRT analysis. As follows: The percentage of phenolic production was calculated % of phenolic Secretion = Number of explants that show phenolic secretion total number explantsx100
