Morphogenetic and physiological effects of LED spectra on the apical buds of Ficus carica var. Black Jack

The use of artificial light sources such as light-emitting diodes (LEDs) has become a prerequisite in tissue culture studies to obtain morphogenetic enhancements on in vitro plants. This technology is essential for developmental enhancements in the growing plant cultures due to its light quality and intensity greatly influencing the in vitro growing explants at a cellular level. The current study investigates the effects of different light-emitting diode (LED) spectra on the growth of apical buds of Ficus carica var. Black Jack. Ficus carica, commonly known as figs is rich in vitamins, minerals, and phytochemicals capable of treating microbial infections and gastric, inflammatory, and cardiac disorders. Apical buds of Ficus carica var. Black Jack, presented morphogenetic changes when grown under six different LED spectra. The highest multiple shoots (1.80 per growing explant) and healthy growing cultures were observed under the blue + red LED spectrum. Wound-induced callus formation was observed on apical buds grown under green LED spectrum and discolouration of the growing shoots were observed on the cultures grown under far-red LED spectrum. Multiple shoots obtained from the blue + red LED treatment were rooted using 8 µM indole-3-acetic acid (IAA), and the rooted plantlets were successfully acclimatised. Compared with the other monochromatic LEDs, blue + red proved to be significantly better for producing excellent plant morphogeny. It is apparent that blue and red LED is the most suitable spectra for the healthy development of plants. The findings have confirmed that the combination of blue + red LED can potentially be used for enhancing growth yields of medicinally and commercially important plants.

Establishment of a Novel and Efficient Agrobacterium-Mediated in Planta Transformation System for Passion Fruit (Passiflora edulis)

Passion fruit (Passiflora edulis) is an important fruit crop with high economic value. Genetic engineering plays an important role in crop improvement with desired traits and gene functional studies. The lack of a simple, efficient, and stable transformation system for passion fruit has greatly limited gene functional studies. In this study, a simple and efficient Agrobacterium-mediated in planta transformation system for passion fruit was established, using Agrobacterium virulent strain EHA105 harboring the binary vectors pCAMBIA1301 and pCAMBIA1302 with GUS and GFP reporter genes. The system requires less time and labor costs than conventional transformation systems, and no additional phytohormones and sterile conditions are required. Regeneration efficiency of 86% and transformation efficiency of 29% were achieved, when the wounds were wrapped with Parafilm and the plants were kept in darkness for 15 days. Approximately 75% of the regenerated plants had a single shoot and 26% multiple shoots. The transformation was confirmed at the DNA and RNA levels as well as by GUS staining and GFP fluorescent measurements. The developed protocol will contribute to the genetic improvement of passion fruit breeding.

Thiol Compounds, Pre-Conditioning and Orientation of Explants – Important Factors Affecting Regeneration from Cotyledons of Legume Crop Sesbania Aculeata

Sesbania aculeata is a multipurpose legume crop grown primarily for green manuring in the rice-based cropping system. Besides this, it is an industrial crop and is also used as food in many parts of the world. The present work reports for optimization of various parameters (growth medium, plant growth regulators, pre-conditioning, orientation of explant, and presence of thiol compounds) affecting in vitro regeneration using mature cotyledon explants. The 5-day-old mature cotyledon explants excised from seedlings grown on Murashige and Skoog (MS) salts and Gamborg (B5) vitamins medium containing 15 μM 6-benzylaminopurine were cultured with its adaxial side facing on medium containing 2.5 μM 6-benzylaminopurine and 50 mg/L thiourea and produced multiple shoots (7 ‒ 8) in 100% cultures within 28 days. Healthy shoots were rooted on half-strength Murashige and Skoog (MS) salts and full-strength vitamins medium augmented with 2.5 μM indole-3-butyric acid.

Shoot Proliferation, Callus Induction And Plant Regeneration In Tripsacum Laxum Nash

The peduncles of Tripsacum laxum Nash were used as explants to induce axillary shoots. Multiple shoots were proliferated on Murashige and Skoog (MS) medium to establish, for the first time, efficient shoot proliferation and plant in vitro regeneration systems. Optimal shoot proliferation medium was MS with 3.0 mg/L 6-benzyladenine (BA) and 0.2 mg/L α-naphthaleneacetic acid (NAA), resulting in a shoot proliferation coefficient of 11.0 within 45 d. Optimal rooting medium was MS with 0.1 mg/L NAA and/or 0.1 mg/L indole-3-butyric acid (IBA), inducing 100% root formation from shoots within 30 d. When young roots, leaf sheaths and shoot bases were used as explants, MS medium with 1.0 mg/L thidiazuron (TDZ) and 0.2 mg/L BA induced most shoots, with the least callus. Shoot bases induced beige-white callus and shoots directly on MS medium with 1.0 mg/L TDZ and 0.2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D), while leaf sheaths induced beige-white callus and shoots directly on MS medium with 1.0 mg/L TDZ and 0.2 mg/L BA. Rooted plantlets showed 99.3% survival when transplanted into a substrate of vermiculite: peat soil (1:3, v/v).

STUDYING THE EFFECT OF A COMBINATION OF PLANT GROWTH REGULATOR ON MULTIPLE SHOOTS AND ROOTING IN GmCHI TRANSGENIC T. PANICULATUM

Thổ nhân sâm (Talinum paniculatum) là loại cây thân thảo được biết đến với giá trị dược liệu cao. Cây Thổ nhân sâm chuyển gen GmCHI ở thế hệ T1 có hàm lượng flavonoid tổng số tăng 7,4 lần và 4,8 lần so với cây đối chứng không chuyển gen. Để tăng thu nhận flavonoid từ sinh khối của các cây Thổ nhân sâm chuyển gen ở thế hệ T1 có hàm lượng flavonoid cao thì việc nghiên cứu hiệu quả tạo đa chồi và ra rễ ở cây chuyển gen là cần thiết. Trong bài báo này, chúng tôi trình bày kết quả nghiên cứu ảnh hưởng của tổ hợp các chất kích thích sinh trưởng lên sự phát sinh chồi và ra rễ ở cây Thổ nhân sâm chuyển gen GmCHI. Đoạn thân mang mắt chồi bên là vật liệu thích hợp để tạo đa chồi khi sử dụng môi trường MS cơ bản bổ sung kinetin. Bổ sung 2,0 mg/l BAP cho hiệu quả đa chồi cao hơn so với bổ sung 1,5 mg/l kinetin hay kết hợp 1,5 mg/l kinetin với 0,8 mg/l IBA. Sử dụng 0,5 mg/l IAA có khả năng tạo rễ tốt hơn so với kết hợp 0,5 mg/l IAA với kinetin.

Effect of Nitrogen on in Vitro Propagation of Endangered Medicinal Plant: Swertia Chirayita Roxb. Ex Flaming

The effect of nitrogen was investigated on the organogenesis of Swertia chirayita (Gentianaceae) to overcome the challenges related to its cultivation. The best callogenic response was observed on root explants inoculated onto MS medium supplemented with BAP (2.0 mg/l) along with 2,4-D (0.5 mg/l) after 35 days of culture. Subsequent transfer of callus for multiplication on the same media composition under complete darkness presents the best results in terms of callus multiplication. Callogenic cultures were subculture onto modified MS medium supplemented with inorganic nitrogen sources, i.e., NH4NO3 (14-56N/l), KNO3 (100-400N/l) with BAP (3.0 mg/l) were observed. Organogenic response (52%) was observed after 8-12 weeks of culturing. The maximum number of the shoot was recorded on MS medium with NH4NO3 (28 N/l), KNO3 (300N/l) with BAP (3.0 mg/l). Moreover, 90% of them were able to regrow when sub-cultured on the same media. Sixteen weeks old multiple shoots were subcultured on MS medium supplemented with different auxins. IAA was proved to be the best hormone rooting purpose. However, the best rooting response regarding the number of roots and an average length of roots was obtained at IAA (1.0 mg/l). Survival of 90% was achieved when rooted plantlets were successfully established in substrate containing sand, vermicompost, and garden soil in equal proportion for hardening and acclimatized.

Effects of Phytohormone and Regulators on Shoot Tip and Nodal Explants for In Vitro Shoot and Root Clonal Propagation of Vitex negundo L.

Medicinal plants are one of the most vital natural resources, but many of them are currently endangered due to habitat loss. Consequently, it is critical to emphasize the importance of using micropropagation techniques for mass propagation of plantlets on a commercial scale, in addition to germplasm conservation and distribution. Nodal explants and shoot tips were expunged from 15 days of the explant by aseptic seedlings, an effective, quick, and better in vitro plant regeneration procedure for Vitex negundo L. has been developed. The recent study was considered to develop an in vitro procedure for the regeneration of V. negundo L., a traditional medicinal plant. Nodal segments and shoot tips were cultivated on MS medium enhanced with numerous plant growth regulators. For multiple shoots and root regeneration, various cytokinins were examined. 6-benzyl-aminopurin (BAP), kinetin (Kin), and 1H-indole-3-butanoic acid (IBA) were all tested as a supplement to Murashige and Skoog (MS) medium including auxin phytohormone, such as Indole acetic acid (IAA) and 1-naphthaleneacetic acid (NAA). The furthermost effective surface sterilization treatment for explants of V. negundo has been found 0.1% HgCl2 for 8 minutes. In all treatments, multiple shoots were collected from shoot tips and nodal segments. In MS media added with 2.0mg/l BAP, the most shoots were seen in V. negundo. Furthermore, V. negundo regeneration shoots rooted effectively in half MS containing 1.0 mg/l IBA. Finally, proliferated plantlets were effectively adapted in soil, where they grew normally without morphological anomalies and had a survival rate of 92 percent.

In Vitro plant regeneration from leaf explants of Tagetes erecta L.

Regeneration of multiple shoots via callus induction and organogenesis was obtained from young leaf explants of the field grown marigold (Tagetes erecta L.). Callus induction and shoot regeneration at various frequencies were observed using different concentrations and combinations of growth regulators. Highest percentage (90%) of callus formation was observed within two weeks on MS medium supplemented with 5.0 mg/l BAP with 2.5 mg/l NAA. The maximum percentage (80%) of shoot bud formation (10±0.5/callus) was obtained from MS medium containing 1.0 mg/l BAP with 0.5 mg/l kinetin. The regenerated shoots developed highest percentages (90%) of roots on half strength MS medium supplemented with 1.0 mg/l IBA. The plantlets when transferred into potsoil 80% survived. Regenerated plants were morphologically uniform with normal leaf shape and growth pattern. Bangladesh J. Sci. Ind. Res.56(2), 69-74, 2021

Sterile Tissue Preparation and Callus Induction of Curcuma longa Linn.

Curcuma longa Linn. (family Zingiberaceae), commonly known as ‘turmeric’, is native to Southeast Asia. Turmeric has been used for color, flavor as a spice in cuisine and employed for treatment of various diseases. The major component in yellow-pigmented fraction of turmeric is curcuminoids. Curcuminoid production in callus of C. longa Linn. is our focus of study. Sterile techniques to obtain germ-free of C. longa Linn. explants were investigated and the results showed that immersing rhizome buds in 70% ethanol for 5 min, followed by 0.10% HgCl2 for 10 min offered approximately 66% survival rate. Multiple shoots were generated from the aseptic rhizome explants cultured on Murashige and Skoog (MS) agar medium fortified with 3.00 µM of 6-Benzylaminopurine (BA) and 0.50 µM of 1-Naphthaleneacetic acid (NAA) at 25 ± 2°C under a photoperiod of 16 h light and 8 h dark. The sterile leaf sheath and root were subsequently used for callus induction which produced various responses when cultured on MS agar medium fortified with different concentrations of 2,4-dichlorophenoxy acetic acid (2, 4-D), Thidiazuron (TDZ) and BA. The highest induction yields of friable callus were obtained from leaf sheath segments cultured on MS agar medium fortified with 0.50 mg/l 2, 4-D which are the conditions proposed for successful production of callus culture of C. longa Linn. Keywords: Callus induction, Curcuma longa Linn., Turmeric, Plant tissue culture

Regeneration of Hemidesmus indicus (L.) R. Br. using in vitro nodes: an alternative method for efficient multiplication of shoots

In vivo nodes of Hemidesmus indicus (L.) R. Br. induced healthy multiple shoots with branching in our earlier studies and thus in the present study, potency of in vitro nodes to regenerate shoots was evaluated. In vitro nodes were excised from eight-week-old shoots and placed in Murashige and Skoog’s (MS) medium fortified with sucrose (3%) and different concentrations of 6-benzyladenine (BA) and kinetin (Kn). After eight weeks, optimum of 5.42 ± 0.36 shoots with 100% response were regenerated in medium supplemented with BA (10 µM) and Kn (5 µM). These healthy shoots were placed in full, half and quarter strengths of liquid MS medium fortified with sucrose (1%) and α-naphthaleneacetic acid (NAA, 1-25 µM) for rooting. Among all the strengths of MS medium, full strength MS medium having 8 µM NAA formed maximum of 3.42 ± 0.55 roots (91.67% response) within four weeks. The protocol is in continuation with earlier study and it was confirmed that a single in vivo nodal explant can regenerate around 385 healthy elongated shoots within 4 months, which will help in mass-propagation of the species.