Regeneration of Shoot from Nodal explants of Cucumis sativus considering different Hormonal concentration (original) (raw)

https://doi.org/10.13140/RG.2.1.4317.1687

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Abstract

DESCRIPTION Regeneration of Shoot from Nodal explants of Cucumis sativus considering different Hormonal concentration Firoz Alom M., Ruhul Amin, Ekhlas Uddin M., Sudhangshu kumar Biswas and Monirul Islam M. Department of Biotechnology and Genetic Engineering, Islamic university, Kushtia, BANGLADESH

In Vitro plant regeneration of Cucumber (Cucumis sativum (L.) from cotyledon and hypocotyl explants

jsrr.in

In vitro plantlet regeneration has been obtained from 15-20days old seedling cotyledon and hypocotyl segments of Cucumis sativus (L.) were examined using various phytoharmons individually and in combination on Murashige and Skoog (MS) semi solid medium supplemented with BAP (1.0-5.0 mg/L) Kn (1.0-5.0 mg/L),IAA (0.5 mg/L)+ BAP (1.0-5.0 mg/L) and IAA (0.5 mg/L)+Kn (1.0-5.0 mg/L) for shoot proliferation IAA (0.5 mg/L)+BAP (3.0 mg/L) was proved to be best for induction of shoots for cotyledon and hypocotyl explants. All regenerated plantlets were rooted on MS medium supplemented with (1.0 mg/L) IAA the regenerated plants grew normally in the green house.

In vitro culture of Cucumis sativus L

Theoretical and Applied Genetics, 1989

The ability to regenerate plants from leaf explants has been tested for three highly inbred cucumber lines (B, G, S), their reciprocal hybrids, F 2 and BC 1 generations. The lines differed from each other in their regenerating ability, which was expressed by the percentage of explants regenerating embryoidal callus and mean number of plantlets per plant. Thus, the lines could be classified as frequently (B), intermediately (G) or occasionally regenerating ones (S). There were no reciprocal cross differences in the regeneration. It was found that the intermediately and intensively regenerating lines contain two pairs of dominant genes responsible for plant regeneration, characterized by complementary and probably additive interaction. The frequently regenerating line differed from the intermediately regenerating in the effect of one gene. It is supposed that the above-mentioned genes belong to three different loci. The ability to regenerate plants from leafexplants had high heritability.

In vitro Plant Regeneration of Two Cucumis melo L. Genotypes Using Different Explant Types and Culture Medium

Ecologia Balkanica, 2019

Development of an efficient in vitro plant regeneration system plays an important role for applying of biotechnological approaches for crop improvement. Therefore, the effect of added of BAP in germination phase on organogenesis of cotyledon and hypocotyl explants of two melon lines was investigated. The seeds germination medium contained three different concentrations of BAP (0.5, 1.0 and 1.5 mg L-1). The rate of plant regeneration was found to depend on genotype, explant type and culture medium. The cotyledons were more effective as explants for organogenesis and subsequent plant elongation than hypocotyls. Combination of 1.0 mg L-1 of BAP in germination medium and 0.5 mg L-1 BAP + 0.5 mg L-1 IAA in next regeneration medium gives the better regeneration answer in two explant types of line 11/9, while in line AGY the most effective was the combination of 1.5 mg L-1 of BAP and 0.5 mg L-1 BAP + 0.5 mg L-1 IAA. The experimental results indicated that pre-treatment with cytokinin BAP (...

Establish of Protocol for Cucumis Melo L. Using Different Hormones Through Plant Tissue Culture Technique

Zenodo (CERN European Organization for Nuclear Research), 2023

Melon (Cucumis melo L.) is economically importance plants in agricultural production in the province of Balochistan, Pakistan. In this study, we developed various regeneration and transformation protocols and we examined the different earlier morphological characters of Cucumis melo on MS medium with different combinations and concentration of plant growth regulators (PGRs) (Auxin, cytokinin, and gibberellin). Result showed that the maximum value of callus was obtained when 0.50mgl-1 of BAP was combined with 0.50mgl-1 of 2,4-D about 52%. The maximum regeneration percentage was obtained by the combination of 1mgl-1 NAA, 0.5 mgl-1 BAP and 0 mgl-1 KIN about 80%. Although, the mean number of shoots were displayed a maximum value in MS medium having the combination of 2.5mgl-1 NAA, 1mgl-1 BAP, and 0mgl-1 KIN about 2.4± 0.3. In addition, when the treatment was treated with 1.5mgl-1 BAP, and 0.5mgl-1 IBA showed a significant response on morphological parameters of studied plants. Similarly, the maximum number of roots, root and shoot length was observed when treated with 2 mgl-1 of IAA and IBA respectively. We assume that statement of the research findings presented here lead to for further studies on in vitro propagation of melon plants.

Determination of Apposite Plant Regeneration Protocol for Several Cucurbits through Direct and Indirect Organogenesis

Horticultural Biotechnology Research, 1970

In the present study, a competent and reproducible practice for the in vitro shoot regeneration of Cucurbita maxima, C. pepo and Cucumis sativus was developed from various explants through direct and indirect organogenesis. In C. maxima, between cotyledon and leaf segment, cotyledon was found to be most responsive for callus induction in MS medium augmented with 0.5 mg L-1 2,4 dichlorophenoxy acetic acid (2,4-D) plus 100 mg L-1 casein hydrolysate and 0.5 mg L-1 2,4-D plus 15% coconut water and for leaf segment it was on MS medium containing 2.5 mg L-1 2,4-D. Comparing the 2 explants it was found that leaf segment was most suitable for callus induction in C. maxima. For massive multiplication of C. pepo mericlones shoot tip and nodal cutting were used. MS medium containing 3.0 mg L-1 6-benzyl aminopurine plus 0.5 mg L-1 gibberellic acid (GA 3) was found most effective for shoot regeneration and 1.0 mg L-1 IBA was found most effective for rooting. In this trait cv. Bulum was more responsive than cv. Rumbo. On the other hand, to generate virus free plantlets of C. sativus, different concentrations of kinetin were used, and 1.5 mg L-1 KIN shown the best performance for primary culture establishment. For shoot multiplication, 1.0 mg L-1 BAP and 2.0 mg L-1 BAP plus 0.5 mg L-1 KIN containing medium shown best result. Subsequently, 2.0 mg L-1 BAP plus 0.5 mg L-1 KIN was best composition for root induction. Our report demonstrated comprehensive protocols and variability in explants, growth regulator response in shoot regeneration potential of in different cucurbit plants.

High Frequency of Multiple Shoots and Plant Regeneration from Different Explants of Cucumis melo L. ‘Flexuosus’: Histological Study and Biochemical Analysis

Within the framework of genetic improvement of a Tunisian Snake-melon (Cucumis melo L.) cultivar by biotechnological methods, we developed a method leading to the regeneration of whole plants by in vitro culturing ofhypocotyl and cotyledon explants, seeds without one cotyledon or with quartile cotyledons, and the embryonic axis, on Murashige and Skoog (MS) medium with different combinations and concentrations of auxin and cytokinin. The percentage of caulogenesis varied with the source of the explant and the composition of the plant growth regulator. The highest percentage of caulogenesis (64.4%) was observed in embryonic axes cultivated on MS with 0.5 mg l-1 2,4-D and 1 mg l-1 BAP. Rooting of buds occurred on MS containing 0.5 mg l-1 NAA. As soon as roots appeared, the plantlets were transferred into pots, and 80% survival was recorded. The origin of shoots was investigated by histological observation. In addition, the uniformity of regenerated plants was checked by polymorphism analysis of six isozymes by starch gel electrophoresis.

Regeneration In Vitro From the Hypocotyl of Cucumis Species Produces Almost Exclusively Diploid Shoots, and Does Not Require Light

HortScience, 2003

Hypocotyl explants of three cultivars of melon (Cucumis melo L.) (cvs. Revigal, Topmark and Kirkagac), and a cucumber (C. sativus L. cv. Taoz) rapidly directly regenerated multiple shoots on Murashige and Skoog medium augmented with 4.4 μm benzyladenine. Regeneration from the hypocotyl resulted in nearly 100% diploid shoots, whereas regeneration from the cotyledons resulted in 40% to 70% polyploid regenerants. Regeneration from cotyledon explants of melon cv. Revigal required light, whereas regeneration from hypocotyl explants of melon cv. Revigal occurred in both light and darkness. Direct regeneration also occurred from the hypocotyl of cucumber cv. Taoz in both light and darkness, even though cotyledonary explants did not regenerate buds or shoots under the same conditions. This is the first report of regeneration from the Cucumis genus producing a fully diploid plant population.

Induction of Multiple Shoots From Shoot Tip Explants of Cucumber (Cucumis Sativus L.)

The present study is focused on induction of multiple shoots from shoot tip explants of cucumber. Shoot tip explants were excised from 11-day-old seedlings and cultured on MS medium fortified with different concentrations of BAP alone (0.5-2.5 mg/l) and/ or in combination with 0.5 mg/l KIN. The highest number of multiple shoot formation was noticed on MS medium supplemented with 1.0 mg/l BAP. The elongated shoots were excised and cultured on MS medium containing different concentrations of IBA (Indole 3-butyric acid) 0.5-2.5 mg/l along with 0.5 mg/l KIN. For rooting, shoots (>2cm) were culture on MS medium containing different concentrations (0.5-2.5 mg/l) of IBA alone and/or in combination 0.5 mg/l KIN. Among the IBA concentrations used, IBA (1.5 mg/l) and KIN (0.5 mg/l) combination was induced maximum percent of rooting (90%) with 7.4 roots/shoot. Rooted plantlets were successfully transferred into plastic cups containing a mixture of soil and sand in the ratio of 1:1 and established in the green house and subsequently acclimatized in the field.

In vitro organogenesis and plant formation in cucumber

Biologia plantarum, 2006

In vitro organogenesis was achieved from callus derived from hypocotyl explants of Cucumis sativus L. cv. Poinsett 76. Calli were induced from hypocotyl explants excised from 7-d-old seedlings grown on Murashige and Skoog (MS) medium containing 87.64 µM sucrose, 0.8 % agar, 3.62 µM 2,4-dichlorophenoxy acetic acid and 2.22 µM 6-benzyladenine (BA). Regeneration of adventitious buds from callus (25 shoots explant-1) was achieved on MS medium supplemented with 8.88 µM BA, 2.5 µM zeatin and 10 % coconut water after two subcultures in the same medium at 30-d interval. Gibberellic acid (1.75 µM) favoured shoot elongation and indole 3-butyric acid (7.36 µM) induced rooting. Rooted plants were hardened and successfully established in soil.

Factors influencing shoot regeneration from cotyledonary explants ofCucumis melo

Plant Cell, Tissue and Organ Culture, 1989

Cotyledonary explants of 4-day-old Cucumis melo cv. 'Hale's Best Jumbo' in vitro seedlings showed maximum initiation of shoot buds when cultured onto a revised Murashige & Skoog medium supplemented with 5/~M indole-3-acetic acid and 5/~M benzylaminopurine and cultured at 25-29°C under low light intensity (5-30#molm-2s-~). Subculture of the shoot buds onto the same medium without auxin and supplemented with 3 #M benzylaminopurine caused the development of shoots from 30% of the buds. The presence of abscisic acid significantly increased the number of explants producing shoot buds. Bud initiation was affected by genotype, seedling age, light intensity, and temperature. Addition of gibberellic acid, thidiazuron or silver nitrate to regeneration medium did not improve either bud initiation or shoot regeneration.

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Regeneration of Shoot from Nodal explants of Cucumis sativus considering Different Hormonal concentration.pdf

International Research Journal of Biological Sciences, 2015

Cucumber (Cucumis sativus), one of the most economically important vegetable (cucurbit) crops, is eaten raw or cooked and is popularly used as salads in south Asia. Considering the nutrition value of Cucumis sativus, the present investigation was undertaken to develop a rapid and efficient in vitro multiplication and regeneration system of this species using in vitro nodal explants. Addition of cytokinin is essential to induce shoot formation from the explants. Of the two cytokinins tested, BAP was more effective than kinetin at concentration of 1.5mg/l yielded best response (87%) on shoot formation. Kinetin at 1.5mg/l showed the highest shoot regeneration frequency (53%). When combination of auxin with IAA (0.5 mg/l) + BAP (1.0 -5.0 mg/l) and IAA (0.5 mg/l) + KN (1.0-5.0 mg/l) for shoots development, greater frequency (70%) were produced at IAA (0.5 mg/l) + BAP (3.0 mg/l) whereas IAA + KN were greater frequency (67%) at 3.0 mg/l KN. For root induction, four concentration of NAA were used. The maximum frequency of root formation (83%) was achieved within 3 week when isolated in vitro raised shoots were cultured on MS medium containing 0.5mg/l NAA.

Exploiting shoot tips as an efficient explant for in vitro regeneration of cucumber (Cucumis sativus L.)

Pure and Applied Biology

A proficient and speedy in vitro regeneration is a prerequisite for genetic modification of plants. The described protocol presents an efficient and practicable procedure for regeneration and multiple shoots induction by using shoot tips as explants for cucumber (Cucumis sativus L.). First, aseptic seedlings were established and shoot tips were excised from those seedlings. The cucumber seedlings used after 7 days of germination were found to be the best source of explants. Theses shoot tips were placed on MS medium supplemented with different concentrations of Benzyl amino purine (BAP) (0.5-2.5mg/L). It was observed that MS medium augmented with 1.5mg/L BAP resulted in maximum shoots formation. By increasing BAP beyond 1.5mg/L, rate of shoots formation was decreased. For rooting purpose, Indole Butyric Acid (IBA) (0.5-2.5mg/L) was used. Shoots greater than 2cm in size were shifted to rooting medium. Best roots development took place on MS medium having 1mg/L IBA. The plantlets with well-developed roots were shifted to peat moss containing pots and gradually acclimated to soil. This in vitro regeneration protocol can be used for genetic transformation of cucumber in future.

In vitro plant regeneration of two cucumber (Cucumis sativum L.) genotypes: Effects of explant types and culture medium

Genetika, 2014

The effect of different phytohormone concentrations on callusogenesis and organogenesis in two cucumber genotypes were studied. It was established that the rate of plant regeneration depends on genotype, explant type and culture medium. Hypocotyls were found to be more responsive than cotyledons in morphogenesis. In vitro planlet-regenerants have been obtained in hypocotyls explants on culture medium with 1.0 and 2.0 mgL-1 BA for cultivar Gergana and in 1.0 and 3.0 mgL-1K-line 15B. Induction of regeneration in cotyledons were established only in cultivar Gergana on culture medium supplemented with 3.0 mgL-1 BA and in combination of 0.5 mgL-1IAA.

Embryogenesis and plant regeneration from anther cultures of Cucumis sativus L

Scientia Horticulturae, 2003

The response of anthers to in vitro culture and effect of growth regulators, temperature pretreatment of anthers have been studied in Calypso and Green Long cultivars of cucumber (Cucumis sativus L.). Direct and callus mediated embryogenesis (embryogenic calli with embryos) was induced in Calypso and Green Long cultivars respectively on B5 medium (Gamborg's medium) supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) and a-naphthaleneacetic acid (NAA) alone as well as 2,4-D in combination with N 6-benzylaminopurine/kinetin/thidiazuron (BAP/KN/TDZ). Optimal embryogenic calli/embryos were induced on B5 medium supplemented with 2.0 mM 2,4-D and 1.0 mM BAP. Temperature pretreatment of flower buds at 4 8C for 0-10 days and also at 32 8C for 1 day were tested and best response was obtained at 4 8C for 2 days. Embryo differentiation was achieved on B5 medium containing 0.09 M sucrose, 0.25 mM NAA and 0.25 mM KN. Embryo maturation was on B5 medium supplemented with 5.0 mM abscisic acid (ABA). Embryos germinated into plantlets on B5 medium containing 0.09 M sucrose. Plantlets were acclimatized in the controlled environment. In each cultivars, the root tips of 24 regenerated plantlets were analyzed for ploidy level, of which 21 and 17 have been haploids in Calypso and Green Long, respectively.

Somatic Embryogenesis and Shoot Regeneration Induced in Cucumber Leaves

Pakistan Journal of Botany

Commercial cucumber cultivars were explored for embryogenesis and plant regeneration induced in somatic tissues on plant growth regulators (PGRs). Maximum callus induction 94.16% and 76% was observed in leaf disc explants on MS medium supplemented with 2,4-D (2 mgL -1), NAA and BAP (1.5 mgL -1 , each), respectively. Seed cotyledon explants induced maximum calli (77%) on 4.0 + 0.75 mgL -1 (BAP + NAA, respectively). Calli induced in leaf disc on the highest level of 2,4-D (5 mgL -1) yielded the highest embryo formation (23%) whereas calli induced on BAP and BAP + NAA (5 + 1 mgL -1) regenerated into 14% and 12%, shoots respectively. These shoots were excised and rooted on MSO medium. The plantlets were transplanted in pots and transferred to field after acclimatization. The developed plant material will be morphologically and genetically characterized for homozygosity.

Multiple Shoot Induction from the Shoot Tip Explants of Cucumber (Cucumis sativus L.)

2001

Introduction: Cucumber is an important horticultural crop in India, cultivated primarily for its fruits for slicing and pickling, juice extraction, and for the preparation of traditional Indian medicines. However, the crop is known to be susceptible to abiotic as well as biotic stress (5). Applications of conventional techniques for its genetic improvement are limited because of interspecific incompatibility. A tissue culture method of propagation that could produce numerous plants would be desirable to produce a sufficient number of plants for inbreeding (2). A system suitable for transformation should ideally produce multiple shoots with minimal peripheral callus production so that the transformation of many of these individual buds is possible (1). This paper describes a technique which allows the production of multiple shoots from adventitious buds in cucumber shoot tip explants.

STANDARDIZATION OF CULTURE CONDITIONS FOR EFFICIENT IN VITRO FLOWERING AND REGENERATION OF CUCUMIS ANGURIA L.

An attempt was made to produce flowering and regenerate gherkin (Cucumis anguria L.) under in vitro conditions using internodal explants. The explants obtained from the seedlings were cultured on MS medium with various combinations of 2,4-D and NAA to obtain callus. The results showed that the rate of callus initiation and formation was high in the medium containing 3.0 mg L -1 2,4-D and 3.0 mg L -1 NAA. Furthermore, the calli were subcultured to a medium containing KN (1.0 mg L -1 ) and IAA (0.5 mg L -1 ) was optimum for the induction of highest number of multiple shoot formation. To study the effects of GA 3 and sucrose on in vitro flowering, the shoots were subcultured on different levels of GA 3 and sucrose separately. It was observed that MS medium fortified with 20 mg L -1 promoted healthy plantlets with 80% of the plantlets producing in vitro flowers on the 4 th week.

Regeneration and frequency of tetraploid variants of Cucumis metuliferus are affected by explant induction on semi-solid medium versus the liquid/ membrane system

Plant Cell Reports, 1998

Ninety-eight percent of Cucumis metuliferus (PI 482439) cotyledons regenerated shoot buds after 5 weeks on basal Murashige and Skoog medium amended with 10 µM benzyladenine. Regeneration rates of explants maintained only on agar-gelled medium versus explants induced first on a liquid/membrane system were compared after weekly transfers of tissue from liquid/membrane to agar during the first 5 weeks of regeneration. The number of regenerant buds increased from six per cotyledon (on agar-gelled medium) to nine per cotyledon when explants induced on the liquid/membrane system for 3 or 4 weeks were transferred to agar-gelled medium. Shoot development, rooting and survival in the greenhouse were adequate, regardless of whether regeneration was initiated on the agar or liquid/membrane system. Tetraploid regenerants accounted for 9% of the 391 regenerants screened. Pollen morphology was a reliable screening technique to identify tetraploid plants. The frequency of tetraploid plants varied from 13.5% to 6.9% after 5 weeks of induction on the agar or liquid/membrane system, respectively. This frequency decreased to 1.5% if the explants were transferred from the liquid/membrane system to agar after the first week of induction. Transfers during this period play a critical role in eliminating tetraploid variants.

Direct plant regeneration from cucumber embryonal axis

Biologia Plantarum, 2007

Embryonal axis explants from 2-d-old in vitro germinated seeds were used to induce multiple shoot production. The combination of 4.44 µM BA and 1.59 µM NAA in MS medium triggered the initiation of adventitious shoot buds. The explants with shoot buds produced maximum number of shoots (10.6 per explant) in MS medium supplemented with 4.44 µM BA and 0.065 mM L-glutamine in three successive transfers. The elongated shoots were rooted on MS medium with 4.92 µM IBA. Rooted plants were transferred to soil with a survival rate of 65 %.