Highest Number Of Shoots Research Articles

SOMATIC EMBRYOGENESIS IN LOCAL COWPEA (Vigna unguiculata L. (WALP) VARIETIES

Organogenesis in vitro was evaluated in explants derived from shoot apices of two local cowpea varieties cultured on Murashige and Skoog (MS) basal medium. To evaluate the response of the cowpea genotypes to in vitro regeneration, seedlings were drived from zygotic embryos excised from surface sterilized seeds after two weeks of culture on MS basal medium + vitamins supplement with 30% sucrose. To regenerate shoots, apices of the In Vitro seedlings with average length of 1cm were excised and cultured on MS supplemented with 0.5, 1.0 or 1.5mg/L 6-Benzylamino purine (BAP) with or without 0.1mg/L α-naphthaleneacetic acid (NAA) for six weeks. Variation was observed in response of the shoot apices from different cowpea cultivars to in vitro regeneration of shoots. The highest shoot number was recorded with Danilla and shoot length was highest in Kanannado, indicating that Danilla responded well to in vitro regeneration. While there were significant differences between these concentrations and other concentrations used in this study on the number of shoots produced, statistically there was no difference between the traetments tested on the in vitro shoot length recorded in this study.Although shoots were succesfully regenerated in this study rooting was not achieved as the explants gradually transformed into calli

Effect of silver nitrate and putrescine on in vitro shoot organogenesis of Polygonum multiflorum

Polygonum multiflorum is a flowering plant that belongs to the family Polygonaceae and it is commonly used for medicinal and ornamental purposes. Few studies have been studied about the regeneration of this species. Therefore, we aimed to develop a suitable protocol for regeneration and subsequent growth of shoots by comparing the silver nitrate (AgNO3) (ethylene inhibitor) and the putrescine (polyamine). Internode explants were cultured on shoot regeneration media (Murashige and Skoog (MS) media containing 2 mg L-1 of 6-benzylaminopurine). To analysis, the effect of AgNO3 and putrescine on shoot regeneration and length, different concentrations (mg L-1) of AgNO3 (0, 1, 5, 7, 10, and 20) and putrescine (0, 10, 30, 50, 100, and 200) were added to the MS media. The result showed that at highest concentration (20 mg L-1) of AgNO3 treatment decrease number of shoots (NOS) (1.4 ± 0.2 mm) and shoot length (9.7 ± 1.6 mm) was observed. Putrescine considerably increased the regeneration efficiency, NOS per explant, and shoot length in all the concentrations when compared to AgNO3 treatment. Among the different concentrations, the highest NOS (2.52 ± 0.2 mm) was obtained in cultures supplemented with 30 mg L-1 putrescine, whereas the further increase in putrescine concentration reduced shoot regeneration. The longest shoots (20.5 ± 1.7 mm) were achieved in cultures supplemented with 200 mg L-1 putrescine. The findings of this study indicate that the addition of putrescine to the media could be suitable for P. multiflorum micropropagation and plant transformation.

Accelerated mono-phasic in vitro mass production of banana propagules and their morpho-cyto-genetic stability assessment

A unique one-step in vitro propagation protocol via concurrent shoot-root development was established in banana cv. Grand Nain. 6-benzyladenine, kinetin, meta-Topolin, thidiazuron (TDZ), and zeatin (0.5—1.5 mg L − 1) individually supplemented in Murashige and Skoog medium were tested for multiple shoot-root regeneration from shoot tip explants. The earliest fresh shoot (∼7 days) and root (∼5 days) initiation, highest number of shoots (∼21) and roots (∼16) with maximum root elongation (16.5 cm) were recorded in 0.5 mg L − 1 TDZ. For all intents and purposes, TDZ (0.5 mg L − 1) was recorded to be the optimum formulation in terms of the highest simultaneous shoot and root formation that was corroborated further by matrix plot analysis. On the other hand, network plot analysis and principal component analysis illustrated the association between plant growth regulators based on their effect on growth attributes under study. Histological studies on shoots, roots, and leaves revealed a consistent growth and development of plantlets under in vitro condition. Cent percent plantlets were survived during acclimatization, and performed best in cocopeat in comparison to other substrata (vermiculite, perlite, vermicompost-soil, soil-sand, and sand only). The cytology-, flow cytometry- and ISSR primer-assisted fidelity assessments of the in vitro-regenerants exhibited numerical chromosomal as well as ploidy stability and monomorphic banding patterns without any polymorphism within the in vitro-regeneranted plantlets and with their mother plant. Based on the comprehensive information developed in this study, this protocol should be of immense importance to facilitate single-step large-scale production of true-to-type planting materials and to explore the possible comparative physico-chemical roles of the plant growth regulators used in this study.

Direct in vitro organogenesis from leaf and internode of Coccinia cordifolia (L.) Cogn.

The current study was carried out to elucidate a reproducible protocol to develop plants directly from leaf and internodes to facilitate the genetic transformation in Coccinia cordifolia (L.) Cogn., a medicinal plant of the cucurbitaceae family. In vitro grown leaf and internodes were used, which were collected for regenerated shoots from field-grown nodal segments that were sterilized by 0.1% HgCl2 treatment for 6 minutes. The nodal segment cultured on BAP, Kn and BAP, NAA combination where 1.5 mg/L BAP solitary supplement and augmented supplement with 0.1 mg/L NAA was most effective as 80% shoots regenerated with 4.0±0.37 and 2.7±0.45 shoots per culture, respectively. Collected leaf and internode responded 90% at 1.5 mg/L BAP + 0.1 mg/L NAA fortified full-strength MS medium. The highest number of shoots also regenerated in the same combination which were 8.1 ± 0.30 and 10.2±0.40, respectively and found internode as the best explant for direct organogenesis. For root induction, half strength of MS medium supplemented with 0.1 mg/L IBA was found most effective. The highest number of roots regenerated per shoot (6.8±0.10) and root length (2.8±0.20). The successful acclimatization of the in vitro (80%) grown plantlets proved the validity of the developed protocol of using biotechnological techniques for improving the plant.

Efficient micropropagation of Thunbergia coccinea Wall. and genetic homogeneity assessment through RAPD and ISSR markers

Thunbergia coccinea Wall. ex D. Don being a rare, ornamental and medicinal plant of India, is needed to propagate for conserving the germplasm and analyzing its phytochemical compounds in the future. A reliable protocol for direct in vitro propagation using nodal shoot meristem of T. coccinea as explant was standardized. The highest number of shoots per explant (22.17 ± 0.54) with maximum shoot length (2.36 ± 0.28) in cm was obtained in Murashige and Skoog (MS) medium supplemented with 9.70 µM of 6-furfurylaminopurine (Kinetin) and 0.053 µM of α-naphthaleneacetic acid (NAA) combination, among all the different plant growth regulators (PGR’s) and concentrations tested. The aforesaid PGR’s combination was optimum for axillary shoot bud induction and multiplication in T. coccinea. The best rooting was observed on the half-strength MS medium fortified with 2.68 µM NAA with the highest number of roots per shoot (3.75 ± 0.12) and maximum length (5.22 ± 0.32) in cm. All the in vitro raised plantlets were acclimatized in sterile sand and soil mixture (1:1) with a survival rate of 70% on earthen pots under greenhouse conditions. PCR-based RAPD (Random Amplified Polymorphic DNA) and ISSR (Inter-Simple Sequence Repeat) molecular markers were employed to determine the genetic homogeneity amongst the plantlets. Twelve (12) RAPD and nine (9) ISSR primers developed a total of 104 and 91 scorable bands, respectively. The band profiles of micropropagated plantlets were monomorphic to the mother, donor in vivo plant, and similarity values varied from 0.9542–1.000. The dendrogram generated through UPGMA (unweighted pair group method with arithmetic mean) showed 99% similarities amongst all tested plants confirming the genetic uniformity of in vitro raised plants.

In Vitro Propagation of Manihot Esculenta Crantz ‘Pirun 2’

Abstract In Thailand, cassava (Manihot esculenta Crantz ‘Pirun 2’) is an important economic crop. The tissue culture technique is used to produce true-to-type cassava plantlets on a large scale within a short period. Therefore, the objective of this study was to investigate the optimum culture medium for cassava axillary bud in vitro propagation. Stem nodes were treated with a solution of 20% (v/v) commercial bleach (25% w/v NaOCl) for 10 minutes. Then, the sterilized buds were cultured on three different types of culture media: Murashige and Skoog (MS), Driver and Kuniyaki Walnut (DKW), and MS supplemented with NAA and BA. Various parameters were evaluated for their effects on plantlet development (height of shoot and numbers of leaves, roots, and shoots after 1 month). The results showed that the mean (± standard deviation) numbers of leaves (6.67 ± 0.82) and roots (2.50 ± 0.55) were significantly higher when they were cultured on MS medium. In addition, the highest number of shoots (3.67±0.82) was obtained on MS medium supplemented with 0.05 mg/l NAA+1 mg/l BA. The cut shoots of plantlets cultured on MS medium without hormones developed into mature plantlets after 1 month. Thus, the MS medium was the optimum for mature cassava Pirun 2 culture. This protocol would be useful for plantlet induction on MS medium supplemented with NAA and BA hormones to produce more shoots and for transfer to MS medium without rooting hormones. Moreover, in vitro derived plantlets were successfully acclimatized on a peat moss which gave the high percentage of transplanting survival (100%). Keywords: Axillary bud, Cassava, Stem node, Surface sterilization, Tissue culture

In vitro propagation and cytogenetic stability of Tripleurospermum insularum (Asteraceae) – a critically endangered insular endemic species from Turkey

Tripleurospermum insularum (Asteraceae) is a critically endangered (CR) insular endemic species in Turkey and is facing high risk of extinction. Here, a rapid and efficient in vitro propagation protocol using nodal segments obtained from seedling shoots cultured on Murashige and Skoog (MS) basal medium supplemented with different plant growth regulators (PRGs) was developed to conserve T. insularum. Besides, the cytogenetic fidelity of propagated plants was tested with DNA ploidy level using flow cytometry (FCM) as well as chromosome counting. The highest shoot number and length of shoot per explant were achieved in MS medium containing 4.6 µM zeatin (ZEA) and 0.5 µM indole-3-acetic acid (IAA). No variation in DNA ploidy level (2x) and somatic chromosome number (2n = 18) of all propagated plants were observed. In vitro rooting of shoots was achieved at 100% efficiency in the medium supplemented with 2.9 µM IAA. The rooted plantlets were transferred ex vitro with 74% survival. This is the first report of a successfully developed micropropagation protocol for ex situ conservation of T. insularum.

Micropropagation and In Vitro Inflorescence of Pentas lanceolata

In this study, different concentrations of 6-benzyladenine (BA) on in vitro shoot and inflorescence inductions of P. lanceolata were investigated. The in vivo and in vitro floral characteristics of this plant were also compared. Nodal explants of P. lanceolata were cultured vertically with the cut ends inserted into semi-solid Murashige and Skoog (MS) medium supplemented with 0, 0.5, 1, 2, 4, and 8 mg L–1 BA. The results showed that the explants formed the highest numbers of shoots even when cultured in MS basal medium without any addition of BA, while the shoots formed in the explants cultured in MS medium supplemented with 1 mg L–1 BA were the longest. No inflorescence was found in the shoots cultured in MS medium supplemented with 8 mg L–1 BA, while the highest percentage of inflorescence induction was found in the shoots cultured in the medium supplemented with 0.5 mg L–1 BA. The apperances of in vivo and in vitro flowers of P. lanceolata were the same in many aspects except that the number of flower/inflorescence formed was different. In addition, water accumulation was observed only inside the in vitro flowers. Water deposit in the long tubular structure of P. lanceolata flower could cause anther injury, suggesting that flowers developed in vitro may not always produce pollen.

Micropropagation of Feverfew (Tanacetum parthenium) and Quantification of Parthenolide Content in Its Micropropagated and Conventionally Grown Plants

Feverfew (Tanacetum parthenium) is a well-known multi-functional plant with anti-inflammatory, cardiotonic, antiangiogenic, and anticancer effects. The therapeutic value of this plant is due to its phytochemical constitutes, especially parthenolide. Tissue culture techniques have been applied to improve the bioactive components of many herbal plants. Hence, this study, was carried out to establish a protocol for micropropagation of the feverfew plant and to quantify parthenolide content in its micropropagated and conventionally grown plants. To establish an aseptic culture, different concentrations of sodium hypochlorite (NaOCl) were investigated for seed surface sterilization. Besides, the effects of plant growth regulators (PGRs) on the callus induction, shoot organogenesis from callus and in vitro rooting were evaluated. Additionally, the parthenolide yield of the micropropagated and conventionally grown plants was determined by using high-performance liquid chromatography (HPLC). The results showed that surface sterilization of feverfew seeds with 6% NaOCl for 15 min obtained 65.00 ± 2.69% aseptic seeds. Murashige and Skoog (MS) medium supplemented with 0.4 mg/L thidiazuron (TDZ) and 2 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) resulted in 86.00 ± 1.72% callus induction. The highest number of shoots (5.00 ± 0.15) per explant was obtained in the treatment of MS medium supplemented with 5 mg/L zeatin. MS medium fortified with 3 mg/L indole-3-butyric acid (IBA) produced the maximum number of roots per plantlet (8.90 ± 0.35). A total of 90% of the micropropagated plantlets survived when planted in perlite + peat moss (1:1 v/v); the micropropagated plantlets were successfully established in the ex vitro conditions. According to parthenolide analysis, its level was significantly higher in the micropropagated plants than conventionally grown plants. Among different solvents, ethanolic extraction obtained the highest parthenolide content of the feverfew plant. Hence, it can be concluded that micropropagation of feverfew could be applied to produce disease-free planting materials and to improve the parthenolide content of the feverfew plant.

Auxin and cytokinin synergism in micropropagation for mass production of Aloe vera.

Aloe vera [Aloe vera (L.) Burm. f.] is considered a valuable medicinal plant worldwide due to its remarkable beneficial effects on human health. However, challenges in A. vera propagation hinder meeting the increasing demand in the health and beauty sectors. As an alternative method, in vitro propagation is crucial for the mass production of Aloe plants, which is a rapid method as well. Therefore, the present study aimed to establish an efficient micropropagation protocol for A. vera by in vitro optimization of the effect of different plant growth regulators (PGRs). For shoot proliferation, sterilized explants were inoculated on the Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (BAP) and thidiazuron (0.5, 1.0, 2.0, and 4.0 mg/l) in combination with 0.5 mg/l naphthaleneacetic acid (NAA). Subsequently, indole-3-butyric acid (IBA) (1.0, 2.0, and 3.0 mg/l) was used for root induction. It was found that the explants cultured on the MS medium supplemented with 4.0 mg/l BAP + 0.5 mg/l NAA showed the highest percentage of response (90 ± 1.29) for shoot induction within the minimum number of days (5 ± 0.33). The highest number of shoots (2.7 ± 0.36) and length of shoots (4.7 ± 0.42 cm) per explant were also observed with the same concentration of PGRs. However, the highest number of roots (3.2 ± 0.57), length of roots (5.67 ± 0.21 cm), and root induction (80 ± 1.97 %) were noticed within the minimum number of days (11 ± 0.79) on the MS medium supplemented with 1.0 mg/l IBA. Thus, the proposed method is a quick and effective approach for the mass propagation of A. vera with appropriate dosages of auxins and cytokinins, which may allow meeting the increasing commercial demand.

Innovative vine training system as a strategy for adaptation to climate change on Verdicchio

Grape quality is endangered by thermal increase caused by climate change. Hence, the necessity to find adaptation solutions moved research activities in the last few years towards experimenting innovative management techniques. In this trial an innovative training system, called High Cane, was compared with the traditional Guyot. High Cane trained vines showed lower photosynthetically active solar radiation values at the fruiting zone, a higher number of shoots per vine and total yield. At harvest, High Cane proved to have some effects in slowing down ripening, i.e. showing lower sugar content, higher titratable acidity and lower pH. The innovative training system is a candidate to be a valid strategy for the adaptation to the effects of climate change.

EFFICIENT IN VITRO PLANTLET REGENERATION FROM COTYLEDON EXPLANTS IN SUGAR BEET (BETA VULGARIS L.)

An efficient in vitro regeneration technique was established in sugar beet (Beta vulgaris L.) using cotyledon as explants from in vitro raised seedlings. Regarding plant growth regulators added in inoculation media showed that BA in combinations with NAA was most responsive for multiple shooting in which, 90% of shoot emergence from inoculated explants were observed with a medium containing 0.5mg/L BA+0.5mg/L NAA at 30 days of inoculation period. The highest number of shoots per explant was 7.5? 0.56 and the maximum shoot length of 6.5?0.25cmwere observed in this medium inoculation after 90 days. In vitro raised shoots rooted well when they were transferred to MS + 3.0mg/LIBA in which, 70% shoots were rooted in this medium transferring after 30 days. The maximum number of roots per shoot was 5.50?0.46 and the average root length of 5.8 ?0.62 cm were recorded inoculation after 45 days. About6 months were needed from culture initiation to plantlets establishment in natural environment and 55% plantlets were found to be acclimatized and resumed a new growth.

CITOCININA E VEDAÇÃO DE FRASCOS AFETAM A PROLIFERAÇÃO DE GEMAS E O DESENVOLVIMENTO In Vitro DE MICROESTACAS DE Jacaranda cuspidifolia MART.

ABSTRACT Plant growth regulators (PGRs) and types of flasks sealing that allow gas exchange can favor the development of in vitro plant propagation systems. The objective of the present study was to evaluate the eff ects of cytokinin supplementation on the induction of shoot proliferation and the influence of gas exchange on the in vitro development of Jacaranda cuspidifolia Mart microcuttings. Nodal segments were cultured in medium supplemented with diff erent concentrations (0.25; 0.5; 1.0 and 2.0 mg L-1) of 6-benzyladenine (BA). In the control treatment, there was no addition of PGRs. After 30 days of culture, the microcuttings were isolated and transferred to a rooting medium supplemented with 1.0 mg L-1 indole-3-butyric acid. At this stage, the culture fl asks were sealed with rigid polypropylene lids with a porous membrane (1 M; 21 μL L-1 s-1 CO2) or with no membrane (0 M; 14 μL L-1 s-1 CO2). Cytokinin supplementation induced the activation of axillary buds of J. cuspidifolia. The highest numbers of shoots were observed in explants cultivated in the presence of 0.5–1.0 mg L-1 BA. The microcuttings kept in flasks with a higher level of gas exchange (1M) had a higher percentage of rooting and greater root and shoot lengths. Microscopic analysis showed a greater differentiation of leaf tissues in plants kept in flasks with a higher level of gas exchange (1M). These plants also showed greater mesophyll thickness and, consequently, greater leaf blade thickness. The results provide new information for establishing an efficient in vitro propagation system for J. cuspidifolia.

Nutrient Media Optimization for Date Palm Micropropagation (Phoenix dactylifera L.)

An experimental study was conducted to optimise nutritional media for date palm (Phoenix dactylifera L.) micropropagation by employing a shoot tip as an explant on basic Murashige and Skoog (MS) medium with varied Auxin (IAA, NAA) and Cytokinin (Kinetin and 2ip) concentrations for shoot and root multiplication. The findings of date palm regeneration on basal MS medium (Control) without Kinetin and IAA indicated a decreased survival rate (5.0%) and no root or shoot up to the end of the experiment. When the basal MS medium having different Kinetin and IAA concentrations showed an increased rate of survival, i.e. basal MS +0.5 mg/L of Kinetin and 0.0 IAA (8.0%), MS basal+1.0 mg/L of Kinetin and 0.5 IAA (6.6) and basal MS +2.0 mg/L of Kinetin and 1.5 IAA (7.0) and basal MS contains 1.5 mg/L of Kinetin and 1.0 IAA resulted in highest survival rate (8.8%). However, the highest number of shoots (2.2) and a maximum length of shoots (8.8 cm) were recorded on basal MS medium containing 2.0 mg/L of Kinetin and 1.5 IAA. Moreover, the highest number of roots (3.34) and measured the maximum length of roots (3.56 cm) was achieved on basal MS media containing 1.0 NAA and 1.5 mg/L of 2ip.

Pengaruh Zat Pengatur Tumbuh dari Sampah Rumah Tangga sebagai Media Tanam Krisan (Dendranthema grandiflora Tzvelev.) secara In Vitro

This study aims to: 1) obtain growth regulators that are easy to obtain at low prices, because synthetic growth regulators are relatively expensive for novice researchers and are very difficult to obtain; and 2) to find out whether household waste can be used as a plant growth regulator in vitro which has been widely used and has been successful conventionally in several types of plants. The method used in this study was experimental, using a completely randomized design (CRD) with three replications. The media used was MS media by adding growth regulators from household waste with a concentration of (0-3) mg/L. The parameters in this study were the number of shoots and roots. The data obtained were then analyzed qualitatively, because the resulting data was not normal and not homogeneous. The results showed that the highest number of shoots was at a concentration of 2 mg/L organic waste growth regulator, with 5 shoots and 15.7 roots, with 8 days after planting on chrysanthemum growing media.

Propagation of Coccinia cordifolia (L.) Cogn. from shoot tip and nodal segment through micropropagation techniques

The present work was undertaken to develop a reproducible protocol for the micropropagation of an important medicinal plant of the Cucurbitaceae family, Coccinia cordifolia (L.) Cogn., by using shoot tips and nodal segments to overcome the impediment in seed settings and seed germination of conventional reproduction. To develop an efficient protocol, 0.1% HgCl2 treatment for 6 minutes was found effective for surface sterilization of field-grown explants to eliminate microbes and fungus and to get healthy tissues. Throughout the study, different concentrations of auxin, cytokine, and gibberellin were used either alone or in combination as supplemented in MS medium to find suitable conditions and suitable explant (nodal segment). BAP was the best cytokine source as 2.0 mg/l BAP produced 4.0±0.20 and 4.8±0.09 shoots per culture and gained 5.8±0.11 cm and 6.7±0.32 cm length with regeneration rate of 90 and 100 percent in shoot tips and node, respectively. The highest percentage (90%) of regeneration for axillary shoot proliferation was obtained in node explants at MS medium containing 2.0 mg/l BAP + 0.1 mg/l NAA. The highest number of shoots regenerated per culture was 3.0±0.41, with a length of 6.0±0.16 cm. GA3 was also found effective in producing longer shoots with the combination of BAP, but the average shoot number was reduced drastically. Although full-strength MS medium was found to be ideal for shoot regeneration and used in shoots proliferated experiments, half and full-strength of MS medium with auxins supplements of 0.5, 1.0, and 2.0 mg/l either of IBA, NAA, and IAA) were used for roots growth and half strength nutrients supplemented with 0.5 mg/l IBA was found most compelling. The highest number of roots regenerated per shoot was 3.1±0.30, and the average root length was 1.8±0.30 cm. Rooted plantlets were finally transplanted into small plastic pots containing sun sterilized sand, soil and humus (1: 2: 1) to adapt the plantlet in ex vitro environment, and acclimatized plantlets showed 95% survival rate in outdoor condition which proved the effectiveness of using biotechnology to improve plant’s growth rate and mass production.

Impact of BAP on in Vitro Regeneration of Potato (Solanum Tuberosum L.)

The potato (Solanum tuberosum L.) plant is grown in about 150 countries of the world and is considered an important food crop. However, this crop is susceptible to different biotic and abiotic factors, which can affect its crop yield. This vulnerability can be reduced or eliminated by growing potatoes under sterilized conditions. Cytokinins, such as 6-Benzylaminopurine, are proven to show a significant role in the in vitro regulation of plants. In the current study, explants of Kuroda variety were/potato cv. Kuroda were grown using diverse concentrations of 6-Benzylaminopurine (BAP), which displayed varied results. BAP concentration of 0.01 mg/l showed a 10 cm shoot length with 41 shoots having 66.66% regeneration efficiency. Meanwhile, the explant grown in 0.25 mg/l BAP concentration showed 16 cm shoot length with 65 shoots having 83.33% regeneration efficiency. On the other hand, the explants that were grown using 0.05 mg/l and 1 mg/l BAP concentration showed 7 cm and 10 cm shoot length with 35 and 52 shoots having 63.33% and 76.66% regeneration efficiency, respectively. Therefore, it was concluded that 0.25 mg/L of BAP showed the best results with the highest number of shoots and shoot length as well as maximum regeneration efficiency among all the tested concentrations.

In vitro propagation from axillary buds of the endangered tree Dalbergia congestiflora Pittier (Fabaceae).

Dalbergia congestiflora Pittier is a woody plant species grown in Mexico and Central America and widely used as timber wood and medicinal material. Since D. congestiflora is an endangered species, an in-vitro micropropagation technique is needed for mass propagation of D. congestiflora plantlets. Nodal segments of D. congestiflora stem cuttings grown in greenhouse conditions were disinfected with an appropriate protocol and in vitro established on Murashige and Skoog medium (MS) supplemented with 0.05 mg l-1 benzylaminopurine (BA). The explants showed 10% contamination with 90% survival, and the initial shoot was regenerated in 90% of them. Axillary buds of 45-day-old initial shoots were cultured on MS containing BA (0, 0.05, 0.1, 0.5, 1, 1.5 and 2 mg l-1) singly or in combination with α-naphthaleneacetic acid (NAA) (0, 0.1, 0.5 and 1 mg l-1). A higher shoot number (9.6 shoots/explant) was obtained on MS with 1 mg l-1 BA and 0.1 mg l-1 NAA. Rooting was investigated using half-strength MS, 2% sucrose and different concentrations of indole butyric acid (IBA) (0, 0.1, 0.5 and 1 mg l-1). After 30 days of culture, developing shoots were elongated and rooted in culture medium without IBA, with production of 3.2 roots/shoot. Micropropagated plantlets of D. congestiflora were successfully transplanted and acclimatized to a mixture of peat moss and perlite (2 : 1) with 100% relative humidity in greenhouse conditions with 80% survival at 30 days of culture. This micropropagation protocol will contribute to the conservation of D. congestiflora, and assure the mass propagation for sustainable usage of this species.

Metal Akümülatör Bitki olan Brassica nigra L.’nin in vitro Kallus İndüksiyonu ve Sürgün Gelişimi

This study established an efficient in vitro callus formation and plant regeneration protocol for a prevıously reported Cu accumulator, Brassica nigra, black mustard collected from Diyarbakir (Station site). Node explants from 10-day old mature plants were used for callus formation and shoot regeneration. The highest callus formation frequency (100%) was observed on Murasige Skoog (MS) medium containing 0.1 mg/L Benzylaminopurine (BAP) + 0.5 mg/L Naphthylacetic acid (NAA) (MS 2), 0.6 mg/L BAP + 0.2 mg/L NAA (MS 7), the highest shoot regeneration frequency (100%) was achieved on MS medium containing 0.6 mg/L BAP + 0.05 mg/L Indole butyric acid (IBA) (MS 8), 0.2 mg/L IBA + 0.2 mg/L NAA (MS 10) and the highest number of shoots per explant (3,25) was obtained on MS medium supplemented 0.6 mg/L BAP + 0.05 mg/L IBA (MS 8). After root, stem, and leaf formation from explants in MS medium, these plants were transferred to soil and grown in the plant growth room for one month. A dependable and effective shoot regeneration procedure was developed, laying the groundwork for genetic transformation in Brassica nigra.

Organogenesis of Cavendish Banana (Musa acuminata L.) Plant in Various Concentrations of ZPT IAA (Indole Acetic Acid) and BAP (Benzyl Amino Purine) Vitro

Bananas are a horticultural commodity that originates from Southeast Asia and the West Pacific, including Indonesia. Banana cultivation is carried out with the aim of increasing the economic value of the community by cultivating the Cavendish banana species. Cavendish banana (Musa acuminata L.) is one type of banana that is widely exported, and has high economic value. The constraints in the cultivation of this Cavendis banana plant are the small, high-quality tillers produced in large quantities and in a short time. Propagation of seeds with tissue culture technology is one solution in dealing with the problem of providing these seeds. This study aims to determine the effect of ZPT IAA and BAP concentrations on the growth of cavendish banana shoots in vitro. This research was conducted at the Tissue Culture Laboratory of the Faculty of Agriculture, Cokroaminoto University, Palopo, on Jalan Lamaranginang, Batu Pasi Village, Wara Utara District, Palopo City. Starting from January to May 2020. This research was arranged in a completely randomized design (CRD), which consisted of 5 treatments and 3 replications with a total of 15 experimental units. By giving treatment concentrations of P0: Control, P1 (IAA 0.5 mg L-1 and BAP 1 mg L-1), P2 (IAA 1 mg L-1 and BAP 2 mg L-1), P3 (IAA 1.5 mg L-1 and BAP 3 mg L-1), P4 (IAA 2 mg L-1 and BAP 4 mg L-1). With research parameters callus color, callus texture, root emergence time, shoot emergence time, number of shoots, and number of leaves. The results showed that at the concentration with treatment P3: IAA 1.5 mg L-1 and BAP 3 mg L-1 produced the highest number of leaves, namely 3 strands. The fastest root emergence time was 25.33 days and the highest number of shoots was 2.33. While for the time of emergence of treatment shoots P0: Control showed the best results, namely 7 DAS (days after planting). This is because a balanced amount of auxins and cytokinins can affect the growth of banana plants in vitro