Effect of seaweed extracts and plant growth regulators on high-frequency in vitro mass propagation of Lycopersicon esculentum L (tomato) through double cotyledonary nodal explant

An efficient and reproducible two-step in vitro propagation system for tomato (Lycopersicon esculentum) was developed by using the combinations of seaweed biostimulant (Gracilaria edulis and Sargassum wightii) extracts and plant growth regulators (PGRs). Double cotyledonary nodal (DCN) explants of Co-3 cultivar were initially cultured on Murashige and Skoog (MS) and Gamborg’s medium (B5) containing thidiazuron (TDZ) and 6-benzylaminopurine (BA); the best responding cytokinin was tested in combinations with different auxins (NAA, IAA and IBA), and seaweed extracts (G. edulis and S. wightii) of about basal MS medium +10–70% was used for shoot proliferation. The best organogenic culture response was obtained on MS medium fortified with 1.5 mg L^?1 TDZ and 1.5 mg L^?1 IBA. Up to 24 shoots per explants were formed at an optimal duration of exposure to 35 days. Mini shoots of about 3–4 cm were transferred to medium supplemented with MS + iP, MS + zeatin, MS + G. edulis and MS + S. wightii at different concentrations. High frequency of shoot elongation was observed in the medium supplemented with 30% G. edulis (15.2 cm), and profuse rooting was observed in the medium supplemented with 50% S. wightii of about 16.1 cm. Shoot elongation and rooting were observed in the medium supplemented with seaweed extracts. The plantlets were transferred to the plant growth chamber (70% of relative humidity and 9 light cycles) and maintained in it for a week, and then they were transferred to a greenhouse condition. The plant growth chamber to green house transferred plantlets showed an increase in the survival rate from 70 to 85%. Thus a two-step regeneration protocol was developed in this study with a combination of seaweed extracts and PGRs, which provides a basis for the production of transgenics with high frequency and survivability of tomato plants.     

Optimization of somatic embryogenesis protocol in Lycopersicon esculentum L. using plant growth regulators and seaweed extracts

In the present study a simple and efficient somatic embryogenesis system was developed from leaf explants of Lycopersicon esculentum L. The protocol has been developed by using plant growth regulators and seaweed extracts a natural biostimulant. The leaf sections were initially cultured on to leaf embryogenic callus induction medium fortified with various concentration and combinations of 2,4-dichlorophenoxy acetic acid (0.2–1.0 mg L^?1), picloram (0.2–1.0 mg L^?1), and kinetin (0.1–0.5 mg L^?1). The best responding concentration in induction of friable embryogenic callus was tested for the proliferation. The friable cultures were detached from the mother culture and inoculated in three different media supplemented with plant growth regulators, plus 0–25 % Caulerpa scalpelliformis or 0–25 % Gracilaria corticata extracts for embryo development. A twofold increase in maturation and germination of somatic embryos was observed in the media containing seaweed extracts (MSMG2 and MSMG3) than the control (MSMG1). The plantlets transferred from plant growth chamber to greenhouse conditions exhibited higher survival rate (90 %) than directly shifted plantlets.     

The effect of growth regulators on tomato (Lycopersicon esculentum) fertility

The aim of the study was to determine the effect of 2,4 D (2,4dichlorophenoxyacetic acid) used separately and in a mixture with BAP (benzylaminopurine) on fertility and some features of tomato fruit. The plant flowers were immersed only once in water solutions of the growth regulators (2,4 D 0.001 %; 0.005 %; 2,4 D 0.001 % + BAP 0.001 %; 2,4 D 0.005 % + BAP 0.005 %). The mean fruit weight, fruit volume, specific weight, and the jelly weight were determined. The number of seeds collected from the fruit was used as a criterion of fertility estimation. No statistically significant differences were found in the mean fruit weight, fruit volume and specific weight. The fruits derived from the plants which were not exposed to the action of growth regulators were characterized by the smallest amount of jelly while the fruits set under the influence of 0.001 % 2,4 D + 0.001 % BAP had the largest jelly amount. The greatest differentiation was found in fertility which ranged from 7.5 seeds from the fruit derived from the plants treated with 0.005 % 2,4 D, to 75.7 seeds from the non-treated plants’ fruit. The BAP addition caused the limitation of fertility reduction.     

Effect of plant growth regulators on plant regeneration of Dioscorea remotiflora (Kunth) through nodal explants

Dioscorea remotiflora (Kunth) is an important wild plant that produces tuberous roots used as a source of food in the Western part of Mexico. Lack of planting material and inefficiency of traditional methods of propagation are the main constraints for implementing large-scale cultivation. In contrast, tissue culture techniques allow increasing multiplication and rapid production of plant material. In this regard, leaves or nodal segments were incubated on MS, B5 and WPM culture media with different PGRs in order to obtain an efficient micropropagation protocol. Leaves explants were unable to inducing shoots or callus. However, nodal segments produced axillary shoots and/or callus in all culture media. MS containing 2.33???M KIN was the most suitable to inducing shoots; an average of 6.6 shoots per segment for 100?% explants was obtained, which displayed also the greater number of nodes (5.0) and leaves (7.9) per segment. A decrease on shoot proliferation was observed combining BA or KIN with 2,4-D or NAA. However, small brownish callus were induced on 100?% of segments using 2.33???M KIN with 5.37???M 2,4-D or 9.30???M KIN plus 2.69???M NAA. In contrast, by adding 2.69???M NAA, 66.4?% of the nodal segments formed shoots and produced also yellowish friable callus on the base of the shoots. Shoots were easily rooted with 8.28???M IBA (96.9?%), displaying the greatest root and shoot biomass, but maximum number of tuberous roots, and root or tuberous root biomass was produced increasing IBA (20.7???M).     

Effect of liquid seaweed extracts on growth of tomato seedlings (Solanum lycopersicum L.)

Seaweed extracts are used as nutrient supplements, biostimulants, or biofertilizers in agriculture and horticulture to increase plant growth and yield. In this study, we examined the effect of liquid seaweed extracts (LSEs) made from Ulva lactuca, Caulerpa sertularioides, Padina gymnospora, and Sargassum liebmannii as biostimulants on the germination and growth of tomato (Solanum lycopersicum) under laboratory and greenhouse conditions using foliar and soil drench applications of LSEs. We assessed LSEs at different concentrations (0.2, 0.4, and 1.0 %) on germination parameters (percentage, index, mean time, energy, and seedling vigor index) and growth parameters (plumule length, radical length, shoot length, root length, fresh weight, and dry weight) of tomato seedlings. Our results indicate that seeds treated with LSEs of U. lactuca and P. gymnospora at lower concentrations (0.2 %) showed enhanced germination (better response in germination rate associated with lower mean germination time, high germination index and germination energy, and consequently greater seedling vigor and greater plumule and radicle length). Application as a soil drench was found to be more effective in influencing the height of the plant (up to 79 cm) than the foliar spray application (75 cm). Plants receiving LSEs of U. lactuca and P. gymnospora showed increased shoot length, root length, and weight. Furthermore, U. lactuca and P. gymnospora were found to be more successful and better candidates for developing effective biostimulants to improve the growth of tomato plants. This study provides important information on the identification and utilization of Mexican seaweed resources for agriculture and is the first study to report on the uses of these seaweeds as a source of liquid extracts as biostimulants in agriculture.     

Effect of temperature on biomass allocation in tomato (Lycopersicon esculentum)

Temperature may influence dry matter partitioning between fruits and vegetative plant parts either directly or indirectly through its influence on development, flower and/or fruit abortion. The objective of the present work was to investigate whether there is any direct effect of temperature on dry matter partitioning between fruits and vegetative plant parts in tomato. A greenhouse experiment was conducted, with alternating 3-week periods of high (23°C) and low (18°C) temperature setpoint. Dry matter partitioning during these 3-week periods was determined from destructive plant harvests at two levels of fruit pruning (3 and 7 fruits per truss). Indirect temperature effects on dry matter partitioning were excluded by fruit pruning.
On average, the fraction of dry matter distributed to the fruits during a 12-week period, starting with the flowering of the fifth truss (28 days after planting), was 0.53 (3 fruits per truss) and 0.70 (7 fruits per truss). These ratios were also calculated for every 3-week period separately and did not depend on the average temperature (18–24°C) during that period.
It is concluded that dry matter distribution in tomato is not significantly affected by temperature directly, which means that the temperature effect (18–24°C) on the generative sink strength is not much different from the temperature effect on the vegetative sink strength.     

The induced increase of chitinase activity in tomato (Lycopersicon esculentum L) cells

The levels of chitinase activity induced with elicitors in tomato cells have been detected. It was shown that enzymatic activity depended on degree of polymerization and concentration of biotic elicitors.     

Increase in the rate of recombinants in tomato (Lycopersicon esculentum L.) after in vitro regeneration

Summary Modification to the cross-over (C. O.) rate of tomato (Lycopersicon esculentum) was attempted by using in vitro plant regeneration. F1 hybrids with the same genetical homozygous background were compared at two loci: bs-ms32 on chromosome I, and aa-d on chromosome II. For each, the genetic distance separating the two markers was about 20 to 30 map units. One cotyledon of each F2 hybrid seedling was used as in vitro tissue culture material, while the rest of the plantlet was grown as a control. Recombination rates of the selfed progenies from each regenerated and matched control couple were compared. For the first set of markers 59,000 seeds were analysed (5 controls' and 7 regenerated progenies), and for the second, 11,000 (5 controls' and 8 regenerated progenies). There were significant increases in the genetic distance between markers in about half the regenerated individuals. For the first set the increases ranged from 6.07 to 6.91 units out of a control distance of the 19.84 to 25.65, corresponding to lengthenings of 30.59 to 35.29%. For the second set they ranged from 4.92 to 6.04 out of a control distance of 25.05 to 26.57, representing increases of 19.64 to 22.75%. Such a phenomenon can be important either from a fundamental or practical viewpoint, regarding selection efficiency in plants, and potential for gene reassortment.The experiment reported in this paper has been submitted by M. Biglary in partial fulfillment of the requirements for Docteur-Ingénieur degree. (Nov. 1982, Laboratoire d'Amélioration des Plantes, Université Paris-Sud, F-91405 Orsay)     

Highly efficlent plant regeneration from mesophyll protoplasts of Indian field cultivars of tomato (Lycopersicon esculentum)

Three Indian cultivars ofL. esculentum were assessed for shoot regeneration from protoplast-derived calli. Consistent yields of viable protoplasts (>9.0×10⁶ g f.wt.^-1) were obtained from leaflets of 14 days old cultured shoots. Protoplast viability (88–94%) and planting efficiency (55–70%) were recorded for the three cultivars. Up to 71% of the protoplast-derived tissues regenerated shoots.Abbreviations BAP 6-benzylaminopurine - 2,4-d 2,4-dichlorophenoxyacetic acid - g f.wt. gram fresh weight - IAA indoleacetic acid - MS Murashige & Skoog (1962) - NAA -naphthaleneacetic acid - Zea zeatin     

mRNA levels of four tomato (Lycopersicon esculentum Mill. L.) genes related to fluctuating plant and soil water status

Expression of the genes le20, ni3212, le25 and lcyp2 was quantified in relation to soil and plant water status for pot-grown tomato (Lycopersicon esculentum Mill. cv. Ailsa Craig) in a replicated trial in a partially environmentally controlled glasshouse. Leaf water potential, relative water content, stomatal conductance and gravimetric soil water content were measured at 0800,1200 and 1600 h on three days during the imposition of three different watering regimes. Paired leaf samples were analysed for mRNA. Plant-to-plant variability was quantified and partitioned. le20 mRNA was found in all leaves and responded both to short-term diurnal fluctuations in water status and to more severe stress, though with reduced sensitivity to the greatest stresses. An overnight decline in le20 mRNA was independent of soil or plant water status. The pattern of expression for ni3212 was similar though ni3212 mRNA was much more abundant and the overnight drop in expression was not significant. le25 mRNA was only detected where leaf water potential was below ?0.9 MPa and increased linearly with further decline in leaf water potential. lcyp2 was constitutively expressed with a diurnal pattern characterized by a sharp depression at 1600 h but with expression being enhanced by increasing water deficit at all times. The control and significance of the four genes are discussed in relation to the proposed function of their respective gene products.     

Role of plant growth promoting Bacteria (PGPRs) as biocontrol agents of Meloidogyne incognita through improved plant defense of Lycopersicon esculentum

Plant and Soil - As a major plant-derived soil organic carbon (SOC) component, lignin-derived phenolic compounds show varying biogeochemical characteristics compared to plant-derived lipid...     

Inhibition of ethylene action prevents root penetration through compressed media in tomato (Lycopersicon esculentum) seedlings

In the genus Prunus , so far, somatic embryogenesis has not been reported either from cell suspensions or from their protoplast-derived cells. Rhizogenic cell suspensions of Prunus avium L., initiated from adventitious roots developed from cotyledon-derived callus of mature zygotic embryos, have been subcultured for more than one year without losing their morphogenic potential. A yield of 8 × 10⁵ protoplasts ml⁻¹ of packed cells with a viability of 98% has been routinely obtained. Optimum cell division frequency (around 2.5% at day 10 and 4–6% at day 15) occurs in agarose lenses, with Murashige and Skoog (1962. Physiol. Plant 15: 476–497)-based medium supplemented with 5 μ M naphthalene acetic acid, 1 μ M benzyladenine and 0.25 μ M zeatin. Colony formation has been achieved after 35 days with a plating efficiency of 3–4%. Cell suspensions have been initiated from protoplast-derived callus. While the older cell cultures express a rhizogenic response, the younger ones contain early stages of somatic embryo development. Ultrastructural examination confirms the polarization of these structures.     

Acetylcholine causes rooting in leaf explants of in vitro raised tomato (Lycopersicon esculentum Miller) seedlings

The animal neurotransmitter acetylcholine (ACh) induces rooting and promotes secondary root formation in leaf explants of tomato (Lycopersicon esculentum Miller var. Pusa Ruby), cultured in vitro on Murashige and Skoog's medium. The roots originate from the midrib of leaf explants and resemble taproot. ACh at 10(-5) M was found to be the optimum over a wide range of effective concentrations between 10(-7) and 10(-3) M. The breakdown products, choline and acetate were ineffective even at 10(-3) M concentration. ACh appears to have a natural role in tomato rhizogenesis because exogenous application of neostigmine, an inhibitor of ACh hydrolysis, could mimic the effect of ACh. Neostigmine, if applied in combination with ACh, potentiated the ACh effect.     

Regulation of lycopene formation in cell suspension culture of VFNT tomato (Lycopersicon esculentum) by CPTA, growth regulators, sucrose, and temperature

The onium compound 2–(4-chlorophenylthio)-triethylamine(CPTA) was used to increase lycopene formation to levels approximatingthose in field–or glasshousegrown fruit, and then growthregulators, sucrose and temperature were used to regulate lycopeneaccumulation. It was found that the native auxin indole–3–acetic acid (IAA) was substantially more effective than 2,4-dichlorophenoxyaceticacid (2,4–D) in promoting lycopene formation, sucroseinhibited lycopene formation (cell basis), and temperature produceda pattern similar to that observed in the field with a temperatureoptimum between 18 and 26 C. Suggestions for further improvementsin technique are included. Key words: Suspension culture, tomato, lycopene     

In vitro propagation of Albizia odoratissima L.F. (Benth.) from cotyledonary node and leaf nodal explants

Summary Cotyledonary node and leaf nodal explants excised from 14-d-old in vitro-grown seedlings of Albizia odoratissima were cultured on a Murashige and Skoog (MS) medium with different concentrations of 6-benzylaminopurine (BAP), N ⁶-(2-isopentenyl) adenine (2-iP) and kinetin, used either solely or in combinations. The highest frequency for shoot regeneration (82.5%), the maximum number of shoots per explant (6.9), and the maximum shoot length (2.55 cm) were obtained from cotyledonary node explants cultured on a MS medium containing 10 μM BAP and 10 μM 2-iP with 30 gl^−1 sucrose. Successful rooting was achieved by placing the microshoots on MS medium with 25 μM indole-3-butyric acid (IBA) for 24h first, then transferring to the same medium without IBA. Of the various substrates tested, vermiculite was best for plant acclimatization, in which 75% of plants survived.     

Growth of tomato (Lycopersicon esculentum) in response to salicylic acid under water stress

Abstract

Plants of Lycopersicon esculentum L. cv. K-25 were subjected to water stress by withholding water for 10 days at 20 (WS I) and 30 (WS II) days after sowing (DAS). Seedlings were sprayed with double distilled water (DDW) or 10^?5M salicylic acid (SA) at 45 DAS. The water stress at earlier stage of growth (20 day stage) was more inhibitory as compared to the later stage (30 day stage). The plants exposed to water stress exhibited a significant (p<0.05) decline in photosynthetic parameters, membrane stability index (MSI), leaf water potential, activity of nitrate reductase (NR), carbonic anhydrase (CA), chlorophyll and relative water content (RWC). A follow-up treatment with SA protected against the stress generated by water and significantly improved the above parameters. However, proline content and antioxidant enzymes increased under drought as well as under SA treatments.     

Interspecific protoplast fusion for the transfer of atrazine resistance from Solanum nigrum to tomato (Lycopersicon esculentum. L)

TAES Technical Article #23184     

Influence of growth regulators and explant type on in vitro shoot propagation and rooting of red sandal wood (Pterocarpus santalinus L.).

In vitro shoot regeneration in Pterocarpus santalinus L. was achieved when detached cotyledons from in vitro germinated seedlings were cultured on MS medium containing NAA (0.1 mg/L), BA (1 mg/L) and kinetin (1 mg/L). The regenerated shoots rooted on 1/4 strength MS medium with IAA (1 mg/L) and the fully developed plantlets were successfully established in the soil.