Expression of Heat Shock Protein HSP90 in Genomic-DNA of Chickpea (Cicer arietinum L.) Callus by Heat Shock Treatment

This study was able to detect of the expression activity of heat shock proteins HSP90 and heat transcription factors HSFs for the first time in callus cultures of chickpea, Cicer arietinum L., that exposed to abiotic shocks, grown on MS medium supplemented with 1.0 mg L-1 naphthalene acetic acid (NAA) and 2.0 mg L-1 benzyl adenine (BA). Heat shock proteins HSPs were constructed for increase of withstand long-term physical shocks, and production of resistant to heat chickpeas plants, this shock was enhancement of tolerance of chickpea callus to abiotic stresses (high - temperatures). Results enhanced the ability of chickpea callus to abiotic stresses bearing and induce of HSF genes to heat shock proteins HSP90 production quickly to removing denatured proteins, avoid apoptosis, thus, supporting tolerance to the sudden action of these shocks. Expression activity of heat shock genes and transcription factors by determined based on polymerase chain reaction qPCR, that explained the gene activity increasing at shocks intensity increased

Rosmarinic acid production in cell suspension cultures of Ehretia asperula Zollinger & Moritz

Plant cell cultures provide an alternative means for producing secondary compounds in food, cosmetic and pharmaceutical industries. Ehretia asperula Zollinger & Moritzi is used as a traditional medicine for the treatment of liver detoxification, ulcers, tumors, inflammation and enhancing the body's resistance in Vietnam. The study was carried out to select suitable callus line for cell suspension cultures of E. asperula Zollinger & Moritzi and investigate the effects of inoculum size, rotation speed and naphthalene acetic acid (NAA) on the proliferation of cell suspension cultures. In addition, the influence of light intensity on the growth and rosmarinic acid (RA) biosynthesis of cell suspension was also surveyed. After 4 weeks of culture, the white to pale yellow friable callus expanded significantly with a fresh weight (FW) of 0.788 g and a high RA content of 2.062 mg/g FW. An appropriate medium for cell proliferation was the liquid B5 medium, which contained 30 g/l glucose, 0.1 mg/l benzyl adenine (BA) and 0.4 mg/l NAA. The results also demonstrated that a 1:20 ratio (w/v) inoculum size, darkness and rotation speed of 90 rpm were the optimal conditions for the proliferation and RA accumulation to 188.217 mg/l in 4 weeks of culture. These findings showed that E. asperula Zollinger & Moritzi cell suspension cultures could be a potential alternative approach for RA production in vitro.

The Pivotal Role of Phenological Stage Enhanced by Plant Origin Preparations in the Process of Rhizogenesis of Rosa ‘Hurdal’ Stem Cuttings

Some biostimulants, including plant origin preparations, act similarly to plant hormones. Moreover, the supplementation of known and unknown rooting cofactors can stimulate rhizogenesis in cuttings. The aim of this research was to assess the response of difficult-to-root and long-rooting stem cuttings of the once-blooming old variety Rosa ‘Hurdal’ to preparations of plant origin. The hypothesis was that plant origin preparations could enhance rooting processes by inhibiting chlorophyll a/b degradation in leaves and postponing leaf senescence, simultaneously increasing the quality of cuttings. The one-bud stem cuttings were made in four phenological stages: (H1) flower buds closed, (H2) open flowers, (H3) just after petal fall, (H4) 7-14 days after petal shedding. They were treated with either standard commercial powder preparations containing 0.4% indolebutyric acid (IBA) or 0.2% naphthalene acetic acid (NAA) as well as with commercial plant origin preparations that this work will henceforth refer to as: Algae Extract, Organic Preparation, and Plant Extract. The cuttings were evaluated after 12 weeks of rooting them in two substrates: peat-perlite and peat-sand (v:v; 1:1). Mean root percentages for both substrates were noted after preparation from stage H1 (74.5 %), H2 (59.5 %), H3 (50.8 %) shoots. The H4 cuttings didn’t root at all and were not considered further. The means for all phenology stages together were the highest by the use of 0.6 % Algae Extract, 0.012 % and 0.02 % Organic Preparation, 0.2 % and 0.4 % Plant Extract. The lowest means were reported for the control cuttings as well as NAA and IBA treatment. Plant origin preparations encouraged growth parameters but did not unequivocally inhibit the decrease of chlorophyll content in the cuttings’ leaves. Rooting percentage depended on the quality of cuttings as well as chlorophyll a/b and soluble protein content in leaves in both rooting substrates.

Regeneration potential of different explants during micropropagation of neem tree (Azadirachta indica A. Juss.)

Azadirachta indica A. Juss., (Neem), a prodigious multipurpose tree, has immense potential to benefit mankind and to protect the environment. In order to investigate the effects of three different explants for its regeneration potential, de embryonated cotyledon, immature zygotic embryo and nodal segments from a 30 year old neem plus tree were used. Half strength MS medium with benzyl amino purine (3 mg/L) and naphthalene acetic acid (0.5 mg/L) and casein hydrolysate (1 g/L) was effective in shoot bud sprouting from both nodes and cotyledons. Half strength MS medium fortified with TDZ (0.2 mg/L) was effective for induction of somatic embryogenesis from zygotic embryos. Shoot buds initiated from the cotyledons produced a maximum number of shoots per explants (4.33) which on further sub culturing induced maximum multiple shoots (15) on half strength MS medium fortified with BAP (1.5 mg/L), NAA (0.5 mg/L) and CH (400 g /L) and the nodal explants induced only 4-5 axillary shoots on further sub culturing. Even though immature zygotic embryos produced more number of somatic embryos per explant (24.97) within a short time (30-45 days), the plantlet conversion was poor (25.52 %). In vitro rooting was observed in half strength MS medium supplemented with IBA (2 mg/L). The regeneration potential of de embryonated cotyledons through a simple regeneration system may be beneficial for efficient mass propagation of selected plus trees of neem.

Effects of plant growth regulators on transient expression of foreign gene in Nicotiana benthamiana L. leaves

Background In the last decades, replicating expression vectors based on plant geminivirus have been widely used for enhancing the efficiency of plant transient expression. By using the replicating expression vector derived from bean yellow dwarf virus and green fluorescent protein as a reporter, we investigated the effects of α-naphthalene acetic acid, gibberellins3, and 6-benzyladenine, as three common plant growth regulators, on the plant biomass and efficiency of transient expression during the process of transient expression in Nicotiana benthamiana L. leaves. Results With the increase of the concentration of α-naphthalene acetic acid, gibberellins3, and 6-benzyladenine (from 0.1 to 1.6 mg/L), the fresh weight, dry weight, and leaf area of the seedlings increased first and then returned to the levels similar to the controls (without chemical treatment). The treatment with α-naphthalene acetic acid at 0.2 and 0.4 mg/L can enhance the level of transient expression of green fluorescent protein, which peaked at 0.4 mg/L α-naphthalene acetic acid and was increased about by 19%, compared to the controls. Gibberellins3 at 0.1–0.4 mg/L can enhance the level of transient expression of green fluorescent protein, which peaked at 0.2 mg/L gibberellins3 and was increased by 25%. However, the application of 6-benzyladenine led to decrease in the level of transient expression of green fluorescent protein. Conclusions The appropriate plant growth regulators at moderate concentration could be beneficial to the expression of foreign genes from the Agrobacterium-mediated transient expression system in plants. Thus, appropriate plant growth regulators could be considered as exogenous components that are applied for the production of recombinant protein by plant-based transient expression systems.

Optimization of selected plant growth regulators on callus induction of Oryza sativa L. var MR 219

MR 219, an indica rice from a cross between MR 137 and MR 151 is a long-grain rice variety of high-yielding, good quality of shape and taste, short maturation as well as resistance to blast and bacterial leaf blight. However, it is regarded as a recalcitrant variety due to its low ratio of callogenesis and regeneration in in vitro culturing. This would be a hindrance to crop improvements such as genetic transformation and other crop improvement methods. The objective of this study was to optimize the concentrations and combinations of plant growth regulators (PGRs) which were 2,4-Dichlorophenoxyacetic acid (2,4-D), Kinetin (KIN) and Naphthalene acetic acid (NAA) on its callus induction that ranging from 0.5 to 15.0 mg/L. The callus was induced from the mature seeds of MR 219 on the Murashige and Skoog (MS) media supplemented with 4.4 g/L of MS powder with vitamins, 30.0 g/L of sucrose and 3.5 g/L of gelrite with the addition of 2,4-D, KIN and NAA in different concentrations and combinations. All media supplemented with 2,4- D had successfully induced the callus and 2.0 mg/L of 2,4-D was the best concentration for the callus induction with a 100% success rate. The addition of 15.0 mg/L of NAA, provided less time taken for callus induction with better callus morphology which resulted in fewer browning problems.

Micropropagation, Characterization, and Conservation of Phytophthora cinnamomi-Tolerant Holm Oak Mature Trees

Holm oak populations have deteriorated drastically due to oak decline syndrome. The first objective of the present study was to investigate the use of axillary budding and somatic embryogenesis (SE) to propagate asymptomatic holm oak genotypes identified in disease hotspots in Spain. Axillary budding was achieved in two out of six tolerant genotypes from the south-western region and in two out of four genotypes from the Mediterranean region. Rooting of shoots cultured on medium supplemented with 3 mg L−1 of indole-3-acetic acid plus 0.1 mg L−1 α-naphthalene acetic acid was achieved, with rates ranging from 8 to 36%. Shoot cultures remained viable after cold storage for 9–12 months; this procedure is therefore suitable for medium-term conservation of holm oak germplasm. SE was induced in two out of the three genotypes tested, by using nodes and shoot tips cultured in medium without plant growth regulators. In vitro cloned progenies of the tolerant genotypes PL-T2 and VA5 inhibited growth of Phytophthora cinnamomi mycelia when exposed to the oomycete in vitro. Significant differences in total phenol contents and in the expression profiles of genes regulating phenylpropanoid biosynthesis were observed between in vitro cultured shoots derived from tolerant trees and cultures established from control genotypes.

An Effective Protocol for Carnation (Dianthus caryophyllus L.) cv. 'Geolei' Explants Sterilization for Successful Callusing and Shoot Regeneration

Carnation is a popular floricultural crop grown widely for its attractive cut flowers. Micro-propagation can be used to create large-scale carnation output. For growth and development, plants require some necessary nutrients as well as growth regulators. Due to the importance of carnation, the present work is carried out using leaf and nodal segments to examine the potential of several plant growth regulators for in vitro callus formation and adventitious shoot regeneration. Explants were sterilized properly with bavistin, sodium hypochlorite and mercuric chloride. The minor contaminated cultures were created by consecutively treating the explants with 0.25% bavistin, 0.50% sodium hypochlorite, and 0.1% mercuric chloride for ten, fifteen, and two minutes. MS media with 2.5 mg/l 2,4-dichlorophenoxy acetic acid (2,4-D) in combination with 0.75 mg/l naphthalene acetic acid (NAA) resulted in the maximum callus induction (90.47%) from leaf explants. Maximum shoots (76.47%) were produced in MS media supplemented with 2.0 mg/l Thidiazuron (TDZ) + 0.25 mg/l NAA. NAA at 1.25 mg/l was most efficient for maximum root induction (83.32%). In the present study, an effective protocol of carnation explants sterilization was optimized for successful callusing and shoot regeneration.

Comparative Effect of Different Treaments in Field Pea (Pisum sativum L.)

A field study was conducted to investigate the various treatments that effect on growth and yield of field pea in RBD (Randomized block design) at Sam Higginbottom University of Agriculture Technology and Science, Prayagraj, Uttar Pradesh during march to May in 2020. The trail consists of 13 treatments combinations. The field pea varieties were used were IPF429. The treatments included T0- control, T1,T2,T3,-Gibberellic acid, T4,T5,T6- Neem leaf extract, T7,T8,T9-ZnSo4, T10T11T12- Naphthalene acetic acid(NAA). All Ten parameters treated with Ga3 shows good results in Yield and shows maximum in field emergence, plant height, Days to 50% flowering, Number of pods, Seed yield per plot, Biological Yield and Harvest index. T0 (Un primed) shows lowest of all treatments. Hence, priming with Gibberellic acid could recommended for pre sowing treatment for field pea.

Effect of Nitrogen and Foliar Application of Naphthalene Acetic Acid (NAA) on Growth and Yield of Baby corn (Zea mays L.)

The experiment entitled effect of nitrogen and foliar application of naphthaleneacetic acid on growth and yield of Baby corn (Zea mays L.) was conducted during the Rabi season of 2020 at the Fodder Production Farm of Livestock Research Station, Sri Venkateswara Veterinary University, Lam Farm, Guntur. Andhra Pradesh. The experiment was laid out in Randomized Block Design with 10treatments and each replicated thrice. Treatments consisted of a combination of three levels of nitrogen (60.0, 90.0, 120.0kg/ha) and three levels of naphthalene acetic acid (20,40,60 ppm). It was found that an application of 120 kg Nitrogen/ha as basal along with foliar spray of 40 ppm naphthalene acetic acid at 25 and 35 days after sowing, was the most suitable treatment for obtaining growth and yield attributes such as plant height (177.60 cm), number of leaves (11.33), plant dry weight (113.58g/plant), chlorophyll content (68.43), Leaf Area Index (8.65) and green fodder yield (20333 kg/ha) with net return (83,701.88 Rs/ha) and B:C ratio (1.68). On the basis of one-year experimentation it is concluded that the application of 120 kg Nitrogen/ha + 40 ppm naphthalene acetic acid, on baby corn improved cob yield by 25 % and was found more productive than the recommended dose of fertilizer.