Influence of gibberellic acid, indole butyric acid, and methanol on morpho-physiological and phytochemical traits in Thymus vulgaris L.

Background: Plant growth regulators [PGRs] have important roles in many processes such as germination, seedling growth, nutrition uptake, morphogenesis, ripening, etc

Objective: This study aimed to evaluate the effect of gibberellic acid, indole butyric acid, and methanol as plant growth regulators on morpho-physiological and phytochemical features in Thymus vulgaris L

Methods: The farm experiment based on a randomized complete block design was performed at 2014. The treatments were included G1M1 [GA3 50 ppm + methanol 20 %], G1M2 [GA3 50 ppm +methanol 40 %], G2M1 [GA3 100 ppm + methanol 20 %], G2M2 [GA3 100 ppm + methanol 40 %], I1M1 [IBA 50 ppm + methanol 20 %], I1M2 [IBA 50 ppm + methanol 40 %], I2M1 [IBA 100 ppm + methanol 20 %], I2M2 [IBA 100 ppm + methanol 40 %], G1I2M2 [GA3 50 ppm + IBA 100 ppm + methanol 40 %], G1I2M2 [GA3 100 ppm + IBA 50 ppm + methanol 40 %], and control [distillate water]

Results: The results showed that the combination of GA3 or IBA along with methanol changed significantly leaf length and width, number of branches, leaf dry weight, stem dry weight, plant dry weight, seed weight, essential oil content and thymol amount. The highest values of the most traits were observed in I1M1 treatment

Conclusion: Methanol combination with one of the GA3 or IBA can improve morpho-physiological and phytochemical traits of thyme [Thymus vulgaris L.]. Thus, the most effective PGRs combination was related to GA3 50 ppm + Methanol 20 % and IBA 50 ppm + Methanol 20 %

Large scale propagation of an exotic edible bamboo, Phyllostachys pubescens Mazel ex H. De Lehale (Moso Bamboo) using seeds.

For ex vitro propagation, seeds of P.pubescens were treated with different concentrations of gibberellic acid (GA3) and germination of seeds was tested both in plastic pots as well as by direct sowing in the nursery beds. Maximum seed germination was achieved when treated with 200 mgL–1 (w/v) GA3. For in vitro propagation, an exposure of nodal explants from in vitro raised seedlings to 0.2 mgL–1 1–phenyl–3–(1,2,3–thiadiazol–5–yl) urea and 1 mgL–1 kinetin supplemented medium for 30 days and thereafter to hormone free Murashige and Skoog basal medium resulted in axillary shoot proliferation. For rooting, in vitro raised shoots were exposed to MS medium containing 2 mgL–1 indole-3-butyric acid for 15 days and then shifted to hormone free medium. On an average, 2.8 shoots were obtained in 75% of the cultures within 4 weeks. Such in vitro raised plants were successfully hardened and shifted to field conditions.

Carotenoid genes transcriptional regulation for astaxanthin accumulation in fresh water unicellular alga Haematococcus pluvialis by gibberellin A3 (GA3).

The fresh water unicellular alga Haematococcus pluvialis is a promising natural source of astaxanthin. The present study investigated the transcriptional expression of carotenoid genes for astaxanthin accumulation in H. pluvialis using real-time fluorescence quantitative PCR (qRT-PCR). With treatments of 20 and 40 mg/L of gibberllin A3 (GA3), five genes ipi-1, ipi-2, psy, pds and bkt2 were up-regulated with different expression profiles. GA20 (20 mg/L of GA3) treatment had a greater effect on transcriptional expression of bkt2 than on ipi-1 ipi-2, psy and pds (>4-fold up-regulation). However, GA40 (40 mg/L of GA3) induced more transcriptional expression of ipi-2, psy and bkt2 than both ipi-1 and pds. The expression of lyc, crtR-B and crtO for astaxanthin biosynthesis was not affected by GA3 in H. piuvialis. In the presence of GA3, astaxanthin biosynthesis genes of ipi-1, pds and bkt2 were up-regulated at transcriptional level, psy at post-transcriptional level, whereas ipi-2 was up-regulated at both levels. The study could potentially lead to a scale application of exogenous GA3 in astaxanthin production with H. pluvialis just like GAs perform in increasing crops production and it would provide new insight about the multifunctional roles of carotenogenesis in response to GA3.

A novel combination of plant growth regulators for in vitro regeneration of complete plantlets of guar [Cyamopsis tetragonoloba (L.) Taub.].

A novel combination of plant growth regulators comprising indole-3-butyric acid (IBA), 6-benzylaminopurine (BA) and gibberellic acid (GA3) in Murashige and Skoog basal medium has been formulated for in vitro induction of both shoot and root in one culture using cotyledonary node explants of guar, (Cyamopsis tetragonoloba). Highest percentages of shoot (92%) and root (80%) induction were obtained in the medium containing (mg/L) 2 IBA, 3 BA and 1 GA3. Shoot regeneration from the cotyledonary node explants was observed after 10-15 days. Regeneration of roots from these shoots occurred after 20 to 25 days. The regenerated plantlets showed successful acclimatization on transfer to soil. This protocol is expected to be helpful in carrying out various in vitro manipulations in this economically and industrially important legume.

Interaction between cochleata and stipule-reduced mutations results in exstipulate hypertrophied leaves in Pisum sativum L.

In the wild type P. sativum, each of the adult plant stem nodes, bears a pair of sessile foliaceous stipules and a petiolated unipinnately compound leaf of 4 to 6 leaflets and 7-9 tendrils. The stipule-reduced (st) and cochleata (coch) single null mutants and coch st double null mutant differ fom the wild type in respectively having sessile stipules of much reduced size, petiolated simple and/or compound leaf-like stipules and no stipules. It is also known that coch leaves are somewhat bigger than st and wild type leaves. Here, pleiotropic phenotype of coch st double mutant was investigated. The morphologies of stipules and leaf were quantified in the field grown plants and microcultured shoots, latter in the presence and absence of gibberellic acid and N-1-naphthylphthalamic acid. The observations showed that as compared to the corresponding plants or shoots of COCH ST (WT) genotype, (a) coch st plants bore leaves in which all the organs were hypertrophied; (b) full complement of leaflets and 3-5 tendrils were formed on leaf; (c) the microcultured coch st shoots were taller despite lower number of nodes, and (d) they also produced leaves in which all the organs were bigger and the ratio of leaflets/tendrils was higher. It was concluded that in coch st double mutant (a) ST function is essential for stipule primordium differentiation, in the absence of COCH function and (b) absence of negative feedback loops between simple stipules and compound leaf for metabolite utilization allows hypertrophied growth in leaves.

Physio-anatomic aspects on the initial growth of Guazuma ulmifolia Lam. seedlings (Sterculiaceae)

This paper aimed to evaluate the initial growth of "mutambo" seedlings in different conditions of light intensity and treatments with gibberellic acid (GA). The seedlings were kept under full sun and 50 percent of shading. Sixty days after the emergence, seedlings were sprayed with: 1) 100 mg L-1 GA3; 2) 200 mg L-1 GA3; 3) control. At the end of the appraisals, seedlings height under 50 percent of shading was compared to the height that were growing under full sun with 200 mg L-1 GA. Stem diameter was lower under shading. Leaf area did not vary among the treatments, but the root system growth was higher under full sun and did not vary among GA levels. The number of stomata, trichomes and epidermal cells on adaxial and abaxial sides was higher under full sun. Total dry masses of leaf and root were highe runder full sun and with 200 mg L-1 GA application. "Mutambo" seedlings presented a higher initial growth under full sun, although with a lower height, diameter, and lenght of the largest root and total dry masses of leaf and root were higher. A concentration with 200 mg L-1 promoted a higher growth.

O objetivo desse trabalho foi avaliar o crescimento inicial das mudas de mutambo em diferentes condições de luminosidade e tratamentos com ácido giberélico (GA). As mudas foram mantidas em sombrite 50 por cento de sombra e a pleno sol e aos 60 dias após a emergência as mudas foram pulverizadas com: 1) ácido giberélico 100 mg L-1; 2) ácido giberélico 200 mg. L-1 e 3) testemunha. Ao final das avaliações a altura sob 50 por cento de sombreamento foi maior comparada com aquelas crescendo a pleno sol com GA 200 mg L-1. O diâmetro de colo foi menor sob sombreamento. A área foliar não variou entre os tratamentos, porém o crescimento do sistema radicular foi maior a pleno sol não variando entre os níveis de GA. O número de estômatos, tricomas e células epidérmicas nas faces adaxial e abaxial foram maiores a pleno sol. As massas secas total, de folha e raiz foi maior a pleno sol e com aplicação deGA 200 mg L-1. As mudas de mutambo apresentaram umcrescimento inicial melhor a pleno sol, embora com menoraltura, o diâmetro, o comprimento da maior raiz e a massaseca total, de folha e de raiz foram maiores. A concentraçãode 200 mg L-1 de GA promoveu maior crescimento.

Seed germination of Cistus creticus L. and Cistus laurifolius L. as influenced by dry-heat, soaking in distilled water and gibberellic acid.

The effects of dry-heat and seed soaking in distilled water or in gibberellic acid on germination performance of Cistus creticus L. and C. laurifolius L. were studied in the present study Germination percentages of two Cistus species were low due to dormancy Soaking in distilled water for 24 hr resulted in 28% germination in C. creticus and 43% in C. laurifolius. Gibberellic acid applications (20, 100 and 250 mg l(-1)) for 24 hr gave 32, 30 and 23% germination, respectively in Cistus creticus and 33, 37 and 28% germination, respectively in C. laurifolius. Dry-heat pretreatments at 50, 80 and 100 degrees C for several times (1-60 min) also significantly increased germination percentage in two species. The highest germination in C. creticus was obtained with dry-heat at 100 degrees C for 1 or 5 min (80% and 83%, respectively) and in C. laurifolius at 100 degrees C for 5 min (87%). A significant increase in germination rate was also achieved under different pretreatments. The physiological dormancy caused by an impermeable seed coat can be overcome by dry-heat pretreatments in two Cistus species.

Impact of cadmium and lead on Catharanthus roseus--a phytoremediation study.

The Madagascar Periwinkle, Catharanthus roseus (L.) G. Don (a valued medicinal plant) was exposed to different concentrations ofheavymetals like, CdCl, and PbCl, with a view to observe their bioaccumulation efficiency. Germination was inhibited by both the heavy metals in the seeds previously imbibed in GA, and KNO, for 24 hr. EC50 (the effective concentration which inhibits root length by 50%) was recorded as 180 microM for CdCl2, and 50 microM for PbCl2. Both alpha-amylase and protease activity were reduced substantially on treatment of seeds with increasing concentrations of CdCl2, and PbCl2. Malondialdehyde (MDA) a product of lipoxigenase (LOX) activity also increased due to the treatment of both CdCl, and PbCl2. When two-months-old plants grown in normal soil were transferred to soils containing increasing amounts of these two heavy metals, senescence of lower leaves and extensive chlorosis were noticed after four days of transfer However, plants gradually acclimatized and after 20 days the chlorophyll content was almost comparable to normal. Plants receiving CdCl2 treatment (250 microg g(-1) and less) became acclimatized after two weeks and started normal growth. But PbCl2 of 432 microg g(-1) and less could not affect the plant growth throughout, after a preliminary shock was erased. In case of CdCl2 treatment, a stunted growth with reduced leaf area, reduced biomass and sterility were recorded after six months, while plants show normal growth and flowering in case of PbCl2 treatment. Total alkaloid was also found to be decreased in the roots of CdCl2 treated plants. No change was observed in case of PbCl2. GA3 treatments to the CdCl2 treated plants show internode elongation and increase in leaf area with relatively elongated leaves and thinning of stem diameter AAS analyses of leaves of treated plants exhibited 5-10% accumulation of cadmium, but there was no accumulation of lead at all.

Effect of cadmium on germination, coleoptile and root growth of barley seeds in the presence of gibberellic acid and kinetin.

Effect of cadmium on barley seeds treated with kinetin and gibberellic acid was investigated. As usual, cadmium has inhibited seed germination, and showed important inhibitory effects on roots and coleoptile growth after germination. In general, increase in cadmium concentration caused a greater inhibition of germination, root and coleoptile growth. The adverse effect of cadmium on root and coleoptile growth was more pronounced than that on germination. While testa was pierced by radicle (an indication of germination), no root or coleoptile development was observed above at concentration of 3-9.5 mM CdCl2xH2O. Low concentrations of cadmium have inhibited the root growth more than it did on coleoptile growth. Treatment of seeds with gibberellic acid and kinetin did not show any significant difference on the effect of cadmium in germination. However, inhibition of coleoptile elongation by cadmium has decreased a very much after kinetin application. The same result, although with lower rates when compared to kinetin, has been obtained for GA3 as well. In addition, the inhibitory effect of cadmium on root growth increased even more after kinetin application. The results have been found statistically significant through the least significant different (LSD) test at levels ofp < 0.05 and p < 0.01.

Influence of pretreatment and desiccation on the germination of Laurus nobilis L. seeds.

The effects of moist stratification, gibberellic acid (GA3) in combination with cold-moist stratification and desiccation on the germination of bay laurel (Laurus nobilis L.) seeds were studied. It was found that the external fleshy pericarp caused dormancy. There was also embryo dormancy and it was broken in seeds lacking pericarp either warm-moist stratification (20 degrees C day/4 degrees C night) or cold-moist stratification (4 degrees C) for 8 and 10 weeks, respectively. Gibberellic acid had little effect on germination percent and germination value of seeds without pericarp when used alone but significantly increased overall germination performance when used with cold-moist stratification. The viability of seeds dropped when the moisture content of the seeds was reduced by desiccation, and the time required to reach the critical moisture levels in the seeds differed in the different desiccation temperatures. The critical minimum seed moisture content for bay laurel was around 15% and the seeds showed the typical behavior of recalcitrant seeds.

Efficient clonal propagation method for Decalepis hamiltonii, an endangered shrub, under the influence of phloroglucinol.

A highly efficient two stage protocol was developed for induction of multiple shoots from single node in vitro shoot tip explants of Decalepis hamiltonii. It was found that phloroglucinol (PG) had synergistic effect on shoot multiplication when added with N6-benzyladenine and gibberellic acid. This protocol uses PG for both multiple shoot induction from nodal explants, elongation of primary shoots and initiation of adventitious shoot formation from primary shoots, which was more in presence of triacontanol (TRIA). Maximum number of shoots per culture was observed on the medium containing N6-benzyladenine (1.1 microM; BA), GA3 (5.8 microM) and PG (800 microM). Sub-culturing of the shoots onto MS medium containing optimum concentration of BA (5.6 microM), PG (200 microM) and TRIA (0.011 microM) produced elongated shoots along with secondary shoot formation. The long shoots were rooted on alpha-naphthalene acetic acid (5.38 microM; NAA) and PG (400 microM) containing medium. The rooted plantlets were hardened and their field survival rate was 80-90%.

Micropropagation of Sapindus mukorossi Gaertn.

Bud break and multiple shoots were induced in apical and axillary meristems derived from one month old seedlings of S. mukorossi on Murashige and Skoog (MS) medium supplemented with benzylamino purine (BAP) 0.4 microM or 0.8 microM alone. A combination of BAP and gibberellic acid (GA3) 0.4 microM and 2.8 microM produced elongated multiple shoots from both types of explants. Excised shoots were rooted on MS medium respectively with indole-3-butyric acid (IBA) 3.4 microM or 2.4 microM. The regenerated plantlets were successfully acclimatized and transferred to soil.

influence of NaCl on germination and Ion contents of two wheat cultivars differing in salt tolerance effect of gilbberellic acid

Effect of 100 mM NaCl on seed germination, plumule and radicle elongation and interaction of GA with NaCl were examined in 2 wheat genotypes differing in salt tolerance. NaCl stress reduced seed germination rate equally in both cultivars. However, NaCl-induced reduction in plumule and radicle elongation in Giza 157 [sensitive cultivar] was greater than in Sakha 8. Salinity stress increased accumulation of Na and Cl while it decreased K accumulation in germinating seeds, and more reduction in Giza 157. Gibberellic acid [105 M] partially alleviated the inhibitory effect of NaCl stress on seed germination, plumule and radicle elongation. Gibberellic acid added to NaCl increased K content and decreased Cl and Na accumulation in germination seeds. The effect of salinity stress on germination, ion accumulation and the interaction of gibberellic acid with salinity was discussed