GhAIL6, a Member of the AP2 Family Gene, Promotes Somatic Embryogenesis in Cotton.

Background Somatic embryogenesis (SE) is the process by which plant somatic cells are cultured in vitro without fertilization to regenerate embryos and develop into intact plants, the difficulty of cotton regeneration has severely limited functional gene research and transgenic breeding. The AP2 family is a relatively large family of transcription factor genes that regulate the process of growth and development, but the role of Aintegumenta-Like6 ( AIL6) in cotton SE has not been reported. Methods The 35S::AIL6:GR vector was constructed and transformed into cotton JH713 by Agrobacterium-mediated method, after 3 years of self-breeding, stable genetic T3 generation positive plants were obtained, identified by Southern, and three lines were selected for the following regeneration experiments.Results The results showed that overexpression of GhAIL6 significantly inhibited the proliferation of callus during the first 30 days, and promoted the embryogenic callus production at about 45 days.Couclusion Our results indicated that GhAIL6 was a key regulator of cotton SE, overexpression of GhAIL6 helped to improve the regeneration efficiency of cotton SE

PENGARUH 2,4- D TERHADAP PENINGKATAN KUALITAS DAN KUANTITAS KALUS YANG DIINDUKSI DENGAN KOMBINASI AUKSIN DAN SITOKININ PADA KULTUR DAUN RUMPUT BENGGALA ( Panicum maximum) VARIETAS TRICHOGLUME

To determine the effect various concentration 2, 4, 6 and 8 mg/l of 2,4- D (2,4- dicloropenoxy acetic acid) on callus induction of Benggala grass variety Trichoglume leaf culture on Murashige and Skoog (MS) standard medium and its organogenesis stimulated using deffent concentration of growth regulator, namely [N6- (2- isopenteny)- adenine] or 2iP (0, 0.2 and 1.0 mg/l ) and (l- naphthalene acetic ent, wacid) or NAA (0, 0.03, 0.16 and 3.0 mg/l) were performed. Percentage of callus were measured and organogenesis from callus were subjected to description analysis . The results showed that callus induction was optimum when 2,4-D was treated at 4 mg/l, friable callus were produced. Percentage production of callus week 4 was 46.2 percent, while using 8 mg/l 2,4 –D the callus production was about 59.7 percent yellowish coloured and more compact callus were produced. Combination of 0 mg/l 2iP ( auxin) + 3mg/l (cytokinin) at 3 weeks showed reseilted 100 % of calluses produced roots, the highest amaunt roots (9,0) was observed in combination 0 mg/l 2iP + 3 mg/l NAA, and the longest root (17,0 mm) was recorded in combination 0 mg/l 2iP + 3 mg/l NAA. Calluses yielded varied from white, yellowish to brown colour.

Soybean Callus—A Potential Source of Tocopherols

In vitro cultures have been used as an effective means to achieve a high level of secondary metabolites in various plants, including soy. In this study, the contents of α-, γ-, and δ- tocopherol were quantified in soybean callus, and their amounts were compared to those of soybeans cultivated using the conventional tillage system with three weed controls (respectively without herbicide and with two variants of herbicide). Soybean callus was produced using Murashige and Skoog 1962 (MS) medium supplemented with 0.1 mg/L 6-Benzylaminopurine (BAP) and 0. 1 mg/L Thidiazuron (TDZ). The highest amount of fresh callus was obtained from soybeans from the conventional tillage system with second weed control (S-metolachlor 960 g/L, imazamox 40 g/L, and propaquizafop 100 g/L) respectively 13,652.4 ± 1177.62 mg. The analyzed tocopherols were in much higher content in soy dry callus than the soybean seeds (5.63 µg/g compared with the 0.35 α-toco in soybean, 47.57 µg/g compared with 18.71 µg/g γ-toco or, 5.56 µg/g compared with 1.74 µg/g β-toco). The highest content of the three analyzed tocopherols was γ -tocopherol, both in callus and soybeans. Furthermore, the data showed that herbicides used in soybean culture significantly influenced both the in vitro callus production and the tocopherol callus content (p ˂ 0.05). Altogether, soybean callus can be an important source of tocopherols, and herbicides significantly influence in vitro callus production and the tocopherol callus content.

Anthocyanin: A Natural Dye Extracted From Hibiscus Sabdariffa (L.) For Textile and Dye Industries

Environmental pollution is one of the major issues faced by all the countries throughout the world. To prevent the environment scarcity and crisis faced in day-to-day life due the increasing chemical industries, usage of chemicals and the effluents processed out after the treatment also consists of some trace elements in them. Hence the extraction of enzymes on natural basis forms an alternative criteria for the production of dye in order to reduce pollution which in turn helps to nourish and protect the environment for future generations. Hibiscus sabdariffa (L) has a rich source of anthocyanins which is further enhanced by callus production and synthesized by increasing the sucrose concentration. Anthocyanin pigments were extracted using acidified ethanol and the dye obtained was screened for GL-MS analysis and its dyeing process in textile industry. The study showed significance properties along with coloring nature on the clothes used. Color of anthocyanin pigment depends on pH maintained and also shows the adaptability towards the nature with varied environmental conditions

Evaluating The Impact of Strigolactone GR24 On Capparis Spinosa L. Callus Production And Phenolic Compound Content

The effect of strigolactones on plants, which has been recently described as a new group of plant hormones, has not been fully characterized. Capparis spinosa L. callus formation using synthetic strigolactone GR24 (0.1 and 0.2 μM) alone or in combination with 1-naphthalene acetic acid (NAA) (2 mg / L) and 6-benzylaminopurine (BAP) (1 mg / L) and its effect on phenolic substance production were evaluated. 2 mg/L NAA+1 mg/L BAP+0.1 μM GR24 was the medium with the highest callus formation (60.3%) and callus fresh weight (120.8 mg). In the phytochemical analysis, the highest total flavonoid and phenolic substance and the highest rutin, quercetin, chlorogenic acid content were found in this application and their amounts increased at various rates compared to the control. Aromatic substances in caper calluses were grouped as sulfur compounds (66.97% -87.53%), aldehydes (4.88% -7.90%), ketones (0.34% -19.3%), hydrocarbons and derivatives (0.56%-5.8%), alcohols (% 1.62-6.08%), others (0.61% -2.37%) and their amounts varied at various hormone applications. When 0.1 μM GR24 was applied alone, the total sulfur compound in callus samples was 87.53% and the dominant substance was found to be methyl isothiocyanate.

PROLIFERATION OF OIL PALM (Elaeis guineensis Jacq.) EMBRYOGENIC CALLUS WITH REPEATED SUBCULTURES IN LIQUID MEDIUM

The availability of high-quality seeds is now a necessity. This is due to a government program to replace oil palm trees in smallholder plantations with high quality seeds. An efficient protocol to produce a large number of embryos is needed. To increase the number of embryogenic callus production, the callus proliferation experiment was carried out through suspension culture. This study aimed to examine the proliferation ability of embryogenic callus from three different oil palm clones, in several repeated subcultures. Liquid MS media added with 1 ppm 2.4-D and 0.1 ppm NAA were used. Embryogenic callus was weighed by 0.1 - 0.2 g, transferred into the liquid media, shaking at 60-80 rpm and 27 ºC for 8 weeks without light. Continues subcultures were repeated up to 7 times. The results showed that the growth rate of embryogenic callus increased in the third and fourth subcultures and then decreased in subsequent subcultures. It also revealed that the entire embryogenic callus from the first subculture up to seventh subculture still has the ability to regenerate into new plants. These results indicate that oil palm embryogenic callus can be proliferated by suspension culture with a limit up to the fourth subculture. Ketersediaan benih kelapa sawit berkualitas saat ini merupakan kebutuhan karena adanya program pemerintah untuk menggantikan tanaman sawit di kebun-kebun petani. Salah satu cara vegetatif yang dapat dilakukan adalah meningkatkan jumlah kalus embriogenik yang dihasilkan melalui pengembangan kultur suspensi. Penelitian ini bertujuan mengkaji kemampuan proliferasi kalus embriogenik dari tiga klon kelapa sawit, pada beberapa kali subkultur yang berulang. Media cair MS dengan penambahan 1 ppm 2,4-D dan 0,1 ppm NAA digunakan untuk memperbanyak 0,1–0,2 g kalus embriogenik, dikocok pada 60-80 rpm dan suhu 27 ºC tanpa cahaya selama 8 minggu. Subkultur berulang dilakukan hingga 7 kali. Hasil percobaan menunjukkan bahwa kemampuan proliferasi kalus dipengaruhi oleh genotip tanaman induk. Rata-rata kalus embriogenik dapat meningkat pada subkultur ke-3 dan ke-4 dan semakin menurun pada subkultur selanjutnya. Kalus embriogenik hasil proliferasi subkultur pertama hingga ke-7 dapat tumbuh menjadi calon tanaman baru. Hasil ini menunjukkan bahwa kalus embriogenik kelapa sawit dapat diperbanyak dengan kultur suspensi pada batas sampai subkultur ke-4.

Callus Production Protocol for Aegle marmelos (L.) Corr.: As a Tool for Extraction of Secondary Metabolites

Background: Aegle marmelos (L.) Corr. is an important medicinal and a fruit tree belongs to the family Rutaceae possessing numerous valuable secondary metabolites. The growing commercial importance for secondary metabolites has led to a great demand in the pharmaceutical industry in recent years. Therefore, an efficient callus production protocol was developed as a tool for extracting valuable secondary metabolites from Aegle marmelos.Methods: For seeds, callus induction was observed under three conditions as with seed coat, after removing seed coat and split into two halves after removing seed coat. For callus multiplication, 1cm2 pieces of initiated calli were used. These explants were established in MS medium supplemented with combinations of 2, 4 D either with BAP or Kinetin. All experiments were arranged according to the completely randomized design (CRD) with 20 replicates at the Tissue Culture Laboratory, Department of Crop Science, Faculty of Agriculture, University of Ruhuna, Sri Lanka, for a period of 1 year. Percentage of fungal and bacterial contaminations and percentage of bleached explants were observed to select the best explant/s. Percentage of responded explants were observed to select the best condition for callus induction and quality of callus. Growth of callus was observed visually by giving a score. Best hormonal combination for callus multiplication was observed as fresh weight and dry weight of callus produced under each treatment.Result: High quality callus with higher growth was observed in all combinations of BAP and 2, 4 D tested: ranging from 0.5 mgL-1 to 1.5 mgL-1 BAP and 1.0 mgL-1 to 2.0 mgL-1 2,4 D in Murashige and Skoog (MS) medium. Initiated calli were further multiplied in MS medium supplemented with 2,4 D combined with either BAP or Kinetin. Highest amount of callus biomass was recorded in the MS medium with 0.5 mgL-1 2, 4 D and 1.0 mgL-1 Kinetin (132.58 gL-1 fresh weight). The optimized protocol could be used to produce higher amount of callus in order to extract secondary metabolites from Aegle marmelos (L).

Effect of Different Strenght of Medium on Germination and Seedling Growth of Tomato and Sucrose Effect on Biomass of Tomato Callus

Bioactive compounds of plant cell culture extracts are currently being looked at for its potential for the development of nutraceutical products. Tomato callus is known to have cytoprotective activity. The availability for explants and its callus production are two important things. This paper reports the effect of different strengths of medium on seed germination and seedling growth of tomato cv. Permata and sucrose effects on their biomass callus. Seeds were grown on 4 kind of germination media. Hypocotyl were taken as explants for callus induction in MS medium supplemented by 2.0 mg/l BA and 0.2 mg/l NAA, 3% sucrose, and subcultured after 20 days on medium with 6 variations of sucrose. The highest values ​​for tomato hypocotyl’s height were obtained from the ½ and ¼ MS, while the highest fresh weight was obtained from the MS 0. Both the fresh weight and dry weight of the callus increased along with increasing sucrose concentration. The explant age and biomass need to be a concern regarding what kind of callus that will be produced. Application of 5% sucrose concentration is recommended for callus production in this study.

PENGARUH JENIS EKSPLAN DAN KOMBINASI ZAT PENGATUR TUMBUH (ZPT) TERHADAP INDUKSI KALUS Begonia bimaensis Undaharta & Ardaka SECARA IN VITRO

Begonia bimaensis Undaharta & Ardaka is a potential ornamental plant, and currently known only from one population in Sumbawa. Propagation programs, both conventional and in vitro culture are necessary to ensure its conservation. The aim of this research is to observe the effects of explant types and plant growth regulator combination (2,4-D and kinetin) in inducing callus from B. bimaensis leaf in vitro. Callus induction was initiated from three parts of leaf explant, namely petiole, leaf base, and leaf lamina. The explants were planted on Murashige & Skoog (MS) medium with addition of 2,4-D and kinetin. Concentrations of 2,4-D were 0, 0.5, and 1 ppm, while kinetin concentrations were 0, 1, and 2 ppm. Each treatment was replicated 10 times. Results showed that leaf base was the best explant used for callus induction. Medium D1K2 (MS + 1 ppm kinetin) showed the fastest time for callus induction that was at 20 days after planting. The highest percentage of callus production (100%) was found on D1K3 (MS + 2ppm kinetin); D2K2 (MS + 0.5ppm 2,4-D + 1 ppm kinetin); D2K3 (MS + 0.5ppm 2,4-D + 2ppm kinetin) and D3K2 (MS + 1ppm 2,4-D + 1ppm kinetin).