Combination Of BAP Research Articles

In Vitro Regeneration of Plectranthus Edulis (Vatke) from Leaf Derived Callus

Plectranthus edulis (Vatke) Agnew is a tuber-bearing food in Ethiopia and used as a traditional medicine to treat different diseases. Vegetative propagated plants are mostly affected by different diseases. The establishment of in vitro plant regeneration system in P. eduils is of potential importance to genetically improve this plant using different biotechnological approaches including genome editing and genetic transformation. Best calli (100 %) were obtained on MS medium supplemented with 1.5 mg/l NAA+1.0 mg/l BAP and 2.0 mg/l NAA+0.5 mg/l BAP. For multiplication, maximum (58.68±6.57) mean numbers of shoots were obtained on 0.5 mg/l BAP in combination with 0.4 mg/l GA 3 . Best rooting (10.15±0.95) was obtained on MS medium supplemented with 2.0 mg/l IBA. After a month, all in vitro rooted and 78 % of microshoots were survived in the glasshouse. This protocol can be used to improve this plant through in vitro screening, genome editing and genetic transformation. Keywords : callus induction, growth regulator, organogenesis, regeneration. DOI : 10.7176/JBAH/9-5-03 Publication date :March 31 st 2019

Optimization of Offshoot Outgrowth in Globe Artichoke Using a Combination of Chemical and Mechanical Treatments

The application of cytokinins is a good tool to promote axillary buds in many species, but plant decapitation or leaf cut-back are also suitable methods. This research aims to establish a strategy for artichoke cutting production using a combination of chemical and mechanical treatments. Two experiments were conducted in Tunisia to investigate the effect of 6-benzylamino purine (BAP) on shoot outgrowth in globe artichoke combined with the leaf cut-back at collar level one week after BAP treatment. The first trial was tested in a spring offshoot nursery and the second one in a field of micro-propagated mother plants grown for two years. Five treatments were tested in both experiments: BAP 0 ppm + no cut-back (T1), BAP 0, 100, 200, and 300 ppm + cut-back (T2, T3, T4, and T5 respectively). Regarding growth aspects, the highest number of offshoots was obtained in T4 for both trials with an increase of 49.2% and 37.8% compared to T2 nursery and field values, respectively. T4 also showed a faster rhythm of shoot emission and the biggest shoot size compared to the other treatments. Significant interactions between BAP treatments and offshoot size were recorded for morphological and weight parameters. Regarding the offshoot mineral composition, relevant differences were observed among BAP treatments; moreover, the higher BAP concentrations induced a significant decrease of NaCl plant uptake. Therefore, the combination of BAP 200 ppm and the leaf cut-back could be a potential method to enlarge the cutting production of globe artichoke also reducing some stressful conditions.

Detection of Somaclonal Variation in Micropropagated and Acclimatized Plantlets of Oryza sativa MRQ 74 from Stem Explants

ABSTRACT: In plant tissue culture system, the excessive supply of plant growth hormones may inhibit further growth and development of the regenerants and sometimes it may induce somaclonal variation. In this study, the optimum concentration of plant growth hormones for in vitro regeneration of Oryza sativa L. cv. MRQ 74 was identified and subsequently the occurrence of somaclonal variation of regenerated plantlets was determined through cytological analysis of in vivo and in vitro grown plantlets. MS media supplemented with 0.1 mg L-1 BAP in combination with 0.1 mg L-1 NAA was found to be the optimum concentration for shoots and roots induction of Oryza sativa L. cv. MRQ 74. Plantlets derived from MS media supplemented with 0.5 mg L-1 2,4-D showed higher survival rate than that of MS media supplemented with 0.1 mg L-1 BAP in combination with 0.1 mg L-1 NAA. The plant height, number of leaf, leaf length and number of seeds per stalk of acclimatized plants were significantly lower (p<0.05) as compared with plants grown in vivo. Cytological analysis of in vivo and in vitro grown Oryza sativa L. cv. MRQ 74 revealed that the presence of 2,4-D in the culture media had significantly decreased the mitotic index (MI) and increased the ploidy level of the cell nuclei. The findings of the present study would be useful for plant breeders and biotechnologist since somaclonal variation provides a useful source of genetic variation for crop productivity and quality improvement.

A suitable vegetative propagation technique for effective fruiting and conservation of a vulnerable medicinal plant, Paris polyphylla smit

Paris polyphylla belongs to the family of Melanthiaceae. The herb has many medicinal properties like anticancer, antibacterial, and antifungal bioactivities. This has led to an unsustainable collection of rhizomes from the wild and pushed the herb to the edge of vulnerability. Therefore, innovative and efficient propagation both in vivo and in vitro is urgent for conservation. The paper illustrates an effective and simple method of sprouting and fruiting formation for conservation of this species through vegetative propagation using various plant growth hormones such as indole-3-butyric acid (IBA), naphthalene acetic acid (NAA), 6-benzylaminopurine (BAP), combination of BAP and NAA at different concentrations (0, 50, 100, 150 mg/l), propagation in the wild, and in the polyhouse using decayed wood, black soil and sand in the ratio 2: 2: 1. Results of the combination of 100 mg/l BAP and 100 mg/l NAA showed the highest percentage of sprouting (93.33%) and fruiting (53.33%). The cultivation in the polyhouse demonstrated a significant percentage of sprouting (93.46%) and fruiting of this species (83.56%).

Rapid and mass multiplication of three peppermint (Mentha piperita L.) Collections through In vitro Propagation

Nodal segments of three peppermint collections Ludhiana (L), Pantnagar (P) and Sarabha (S) were cultured on the MS basal medium, enriched with various combinations of auxins and cytokinins. Maximum establishment was recorded in ‘L’, (84.30%) followed by ‘S’ (77.50%) and in ‘P’ (77.30%) on MS supplemented with BAP. The average number of shoots per explant ranged from 1.0 to 5.6 in all the three collections. In ‘P’ the maximum number of shoots per explant was 5.6 followed by 4.8 in collection ‘S’ and 4.4 in collection ‘L’ on MS media supplemented with different combinations of BAP and Kinetin. The average shoot length ranged from 3.0cm to 9.6cm depending upon the media and genotype used. The maximum shoot length of 9.6cm was recorded in collection ‘P’ followed by 8.8cm in ‘S’ and the 8.5cm in ‘L on the same media. Supplementation of IBA (0.25 to 1.50mg/l) to MS media induced root regeneration in cultures ranging from 47.00 to 88.00 per cent in all the three collections. Addition of IBA (1.00mg/l) into MS media produced the root length maximally in all the three collections (2.90cm in ‘S’, 2.84cm in ‘P’ and 2.76cm in ‘L’). The highest average number of roots per explant (3.60) was recorded in collection ‘S’ and the lowest average number of roots per explant (1.10) was recorded in ‘L’. The number of days taken for root initiation ranged from the highest (21days) in variety ‘L’ to the lowest (9 days) in variety ‘P’ on root regeneration medium. Present study reports an efficient and reproducible protocol for rapid and mass in vitro multiplication of peppermint cultivars.

Effect of plant growth regulator on in-vitro callus induction and shoot proliferation of a natural sweetening crop, Stevia Rebaudiana (Bertoni)

Stevia is the world's only all natural sweetener with zero calories, zero carbohydrates and a zero glycemic index. Harvested from a plant in the daisy family, Slevia provides a truly delicious and healthy alternative to sugar or chemical sweeteners. It is medicinal important plant for diabetic patients. In this study, Surface sterilized leaf explants of stevia rebaudiallu Bertoni were cultivated in MS medium supplemented with the combination of 2 - 4 D together with N-(2-furfurylamino)-1-H-Purine-6-amine (Kinetin) (0.5 + 1.0, 1.0+2.0, 1.5+3.0 rugll respectively) and 2–4 D together with NAA (0.5 +1.0, 1.0+2.0, 1.5 +3.0 mg/l respectively) for the callus induction and for the shoot proliferation MS medium supplemented with different combination of BAP and NAA (i.e. 0.5 + 0.5, 0.5 + 1.0, 0.5 + 1.5, 0.5 + 2.0, 1.0 + 0.5, 1.0 + 1.0, 1.0 + 1.5, 1.0 + 2.0). In the present study, we observed that the leaf explants cultured on concentration of 2 - 4 D with NAA (1.5 + 3.0 mg/I), showed maximum response for callus induction and the explants cultured on 0.5 mg/l BAP + 1.0 mg/I NAA showed maximum shoot proliferation (85%).

In vitro Propagation of Indian Teak (Tectona grandis L.) from leaf explants

Efficient plant regeneration via direct organogenesis was established using leaf explants of Tectona grandis L. were tested for growth and organogenic capacity on MS medium fortified with various concentrations of 6-Benzyl Amino Purine and Kinetin (0.5–3.0mg/L) either singly or in various combinations of IAA/IBA (0.5–3.0 mg/L). The highest shoot regeneration frequency (100%) was obtained from leaf explants on MS medium fortified with (1.5 mg/L) IBA/IAA in combination of (2.0mg/L) BAP. At the end of 6 weeks the regenerated shoots were transferred on the same medium (MS + 1.5 mg/L IAA/IBA + 2.0 mg/L BAP) for further proliferation and elongation. The regenerated shoots were rooted with high frequency (60%) in MS medium supplemented with (1.5 mg/L) IBA when compared to other auxin IAA. The in vitro raised plantlets were successfully established in green house and transplanted to natural conditions with 70% survival.

Effect of nutritional and growth hormonal factors on in vitro regeneration of papaya (Carica papaya L. cv. Red Lady)

An efficient in vitro regeneration system was developed for the hermaphrodite papaya ‘Red Lady’ using nodal explants. Nodal explants were cultured on modified Murashige and Skoog (MS) medium supplemented with 3 6-benzylaminopurine (BAP) concentrations (0.0, 0.5 and 1.0 mgL -1 ) or kinetin (10 mgL -1 ) in combination with 1-naphthaleneacetic acid (NAA) (0.0, 0.1, 0.3, 0.4 and 0.5 mgL -1 ). Media consisted of 0.5 or 1.0 mgL -1 BAP in combination with 0.1 mgL -1 NAA were determined to be the most effective based on the percentage of explants that produced shoots (93.33 and 86.67 % , respectively), mean number of shoots (1.79 and 1.81 shoots, respectively) and shoot height (2.26 and 1.95 mm, respectively). Shoot multiplication and elongation was achieved 30 days after transferring the initiated shoots to full-strength MS supplemented with 0.5 mgL -1 BAP, 0.1 mgL -1 NAA, and 200 – 400 mgL -1 casein hydrolysate (CH). Effect of full and haft-strength MS medium supplemented with different indole-3-butyric acid (IBA) concentrations (0.0 – 4.0 mgL -1 ) was tested for rooting. Adventitious rooting occurred after subculturing excised shoots on a medium containing 2.0 or 2.5 mgL -1 IBA and 300 mgL -1 CH, and plantlets were successfully acclimatised on potting medium consisting of peat moss: perlite (1:1). After 40 days of acclimatisation, plantlets grew normally and vigorously. This protocol can be used for commercial production of ‘Red Lady’ papaya plantlets.

RELİKT ENDEMİK LIQUIDAMBAR ORIENTALIS (Miller )’İN HİPOKOTİL VE KOTİLEDON EKSPLANTLARINDAN ADVENTİF SÜRGÜN OLUŞUMU

In this study, the effect of Plant Growth Regulators (PGRs) [6-Benzylaminopurine (BAP)(0, 1.0, 2.0, 3.0 mg L-1) alone and BAP in combinations with α-Naphthaleneacetic acid (NAA)(0, 0.05, 0.10, 0.15 mg L-1) or Indole-3-butyric acid (IBA)(0, 0.01, 0.05, 0.10 mg L-1)] on direct organogenesis from hypocotyl and cotyledon explants of 4-5-week-old sterile seedlings of Liquidambar orientalis (Oriental sweetgum) were investigated. For organogenesis, the addition of BAP to the medium was required, also the combinations of BAP with NAA responded better in terms of the percentage of shoot-forming explants, the number of shoots per explant and the growth of shoots as compared to combinations of BAP with IBA. The highest average percentage of shoot formation (76.64%) was obtained from hypocotyl explants cultured on Woody Plant Medium (WPM) containing 0.10 mg L-1NAA + 3.0 mg L-1BAP. The highest average shoot number per explant (32.20 shoots/explant) was observed in also hypocotyl explants cultured on WPM supplemented with 0.10 mg L-1IBA + 1.00 mg L-1BAP. The combination of 0.05 mg L-1NAA + 1.0 or 2.0 mg L-1BAP was found to be appropriate for both explants.

Effect of Explant Types and Benzyl Amino Purine Concentrations on the In Vitro Regeneration of Several Local Eggplant Cultivars

Eggplant as a vegetable is generally fresh consumed. Besides having a high nutritive value, eggplant contains solasonine and solamargine, which act as an antibacterial that can inhibit cancer cells, gastrictic and respiratory disorders. Eggplant also provides a unique system to study in vitro regeneration since it can be induced from different explants by different growth regulators and morphogenetic pathways. This study aims to observe the effect of explant types and BAP concentrations on the in vitro regeneration of three eggplant cultivars, Rimbang, Tanteloh and Limao. Leaf and hypocotyl explants were cultured on MS semi solid medium supplemented with BAP and IBA in combination. BAP tested at 1, 2 and 3 mgL-1 while IBA tested at 1 mgL-1. Medium containing 1 mgL-1 IBA used for rooting of regenerated shoot. The experiment was arranged in completely randomized design with five replicates. Each replicate consisted of five leaves and hypocotyl explants. Cultures were incubated at 25±10C, 65% humidity, and 16 hours per day photoperiod. Observation was done for 8 weeks of culture. The result indicated that leaf explants showed shoot initiation within 5 – 9 days of cultured, while hypocotyl explants showed the response within 7 – 12 days of cultured. Shoots formation preceded by swollen on the explants surface followed by emerging light green shoot. Leaf explants cultured on medium supplemented with 3 mgL-1 BAP + 1 mgL-1 IBA showed the maximum number of shoots regeneration while hypocotyl explants showed less of shoots in all medium. Similarly, leaf explants cultured on medium supplemented with 3 mgL-1 BAP + 1 mgL-1 IBA showed the maximum number of roots. Among the cultivars, Rimbang showed the best response than other cultivars.

In vitro Mass Propagation from Shoot Tip of Dendrobium Red Bull - An Endangered Epiphytic Orchid

An experiment was carried out to optimize the doses and combinations of growth regulators for in vitro propagation of Dendrobium Red Bull through shoot tip culture. The experiment was laid out according to completely randomized design with 15 treatments and three replications. For multiple shoot induction, newly grown shoot buds (keikies) were cultured on MS supplemented with various combinations of NAA and BAP. The regenerated plants were transferred to ex vitro in different substrates and acclimatized. Best response for the shoot length (21.19 mm) was obtained from the medium supplemented with 3.0 mg/l BAP with 1.5 mg/l NAA. Similarly maximum number of leaves was observed from the same combination at all the recorded days except 60 days, whereas the maximum shoot number (7.66) was obtained from the medium containing 3.0 mg/l BAP + 1.0 mg/l NAA. The shoots were then transferred to root inducing medium and well rooted plants were transferred to different substrates. Survivability of the regenerated plants varied from 60 to 92%. The substrate containing cocodust showed maximum survivability (92%) whereas minimum (60%) was observed from the substrate containing course sand. BAP and NAA proved to be good growth regulators for in vitro multiplication of Dendrobium.
 Plant Tissue Cult. & Biotech. 28(2): 161-169, 2018 (December)

Growth regulators affect the dry weight production, carvacrol and thymol content of Lippia gracilis Schauer

The essential oil from Lippia gracilis Schauer (Verbenaceae) leaves are rich in thymol and carvacrol with antibactericidal and antinflammatory action. The aim of the present study was to evaluate the effect of growth regulators and their combinations on shoot proliferation, growth, and volatile fraction analysis of Lippia gracilis in vitro. In the first experiment, five different concentrations of 6-benzylaminopurine (BAP) were used: 0.0, 2.22, 3.33, 4.44, and 5.55 μM and three of naphthaleneacetic acid (NAA): 0.0, 2.68, and 5.36 μM, in a factorial design. In the second, the treatments consisted in combination of BAP + TDZ at concentration (μM): 0 + 0, 1.11 + 1.13, 1.33 + 0.91, 0.88 + 1.36, 0.43 + 1.81, and 1.78 + 0.46. At 30 days of culture in ½ Murashige and Skoog (MS) medium, the growth and volatile constituents were evaluated. Growth regulators significantly influenced in vitro growth of L. gracilis. Growth data for the first experiment were predicted from the three-dimensional (3D) response surface of the different variables analyzed. In the second experiment, the presence of growth regulators reduced shoots and root length in all combinations. The combinations among regulators (BAP and TDZ) stimulated the number of shoots per explant, but BAP and NAA induced higher shoot number. Variation in the number, content, and profile of volatile compounds were also observed under the influence of growth regulators. The major constituents ρ-cimene, γ-terpinene, thymol, carvacrol, and E-caryophyllene were identified, independent of the experimental conditions. However, the use of growth regulators significantly reduced carvacrol and thymol levels.

<i>In Vitro</i> Propagation of a Sporadic Medicinal Plant <i>Cleome monophylla</i> L

A sporadic medicinal plant Cleome monophylla L. comes under the family Capparidaceae selected for the in vitro regeneration. The leaf explants showed differential responses to various combination and concentrations of BAP, TDZ and NAA and produced yellow to green friable and compact and dense calli. The maximum amount of callus per explants was produced on MS medium with 4.44 µM of BAP, 1.16 µM of TDZ and 1.135 µM of NAA combination and concentration gave the 84% response and produced green compact calli, 19.98 ± 0.03 shoots from the callus and the maximum mean number of ischolar_mainlets per culture was also noted (15 ± 0.57) in this combination with MS medium of half strength. The in vitro developed entire plantlets were fruitfully acclimatized to thermocol cups and earthen pots.

Adaptation of acid-tolerant Leucaena leucocephala cv Tarramba Calluses after gamma irradiation on regenerative media

The aim of this study to measure adaptation of callus Lamtoro (Leucaena leucocephala cv. Tarramba) after gamma irradiation on regenerative media. Lamtoro plant breeding in the form of callus with gamma ray irradiation is required for the development of a new plant-tolerant strain line with a gene mutation approach in tissue culture. The experimental design in this study was a complete randomized design (CRD). There were 13 treatments consitsing of control (M0). 6-benzyl amino purine (BAP) media (M1-M4). kinetin media (M5-M8). and combination of BAP and kinetin media (M9-M12) during 8 weeks. The measured variables were viability of callus. the addition of diameter and height callus. colour and texture callus. Data were tested using analysis of variance (ANOVA) and continued with Duncan test if there were a significant difference among treatment. Data of viability colour and texture used descriptive analysis. The data showed that formulation media M12 (combination 1 mg L−1 BAP and 1 mg L−1 kinetin) was significantly different (P < 0.05) in 8 week after plant (WAP). It was concluded that the best regenerative media for adaptation of callus lamtoro was combination of BAP and kinetin media.

In vitro plant regeneration of Paulownia tomentosa (Thunb.) Steud. from shoot tip and leaf segment

An efficient protocol was developed for in vitro mass propagation of Paulownia tomentosa (Thunb.) Steud. using shoot tip and leaf segment explants from field grown plant. Different concentrations and combinations of BAP, Kn, zeatin and NAA were used for multiple shoot regeneration. Among two types of explants, leaf segment produced the highest number of shoots per explant (12 ± 0.4) when they were cultured on MS supplemented with 3.0 mg/l Kn and 0.5 mg/l NAA. Addition of 10% CW to above mentioned medium increased the number of shoots (18) per culture. Shoot tip explants also produced multiple shoots in the same medium, but their performance was not good as leaf segment explants. For shoot elongation, 100 mg/l urea was more effective when added with best shoot induction medium. Shoots rooted well in halfstrength MS supplemented with 2.0 mg/l NAA, within 12 - 15 days. Regenerated plantlets were successfully acclimatized and established in poly bag containing a mixture of soil and compost in 2:1 ratio. About 90% plantlets survived under open field conditions.

Micropropagation of yardlong bean (Vigna unguiculata (L.) Walsp. ssp. Sesquipedalis L. Verdc.) through in vitro culture

An in vitro propagation protocol has been established for yardlong bean (Vigna unguiculata ssp. sesquipedalis L. Walp. Verdc.) using shoot tip, nodal and leaf segment explants of the superior plant type obtained from crossing of two selected parental lines. Explants were cultured on MS medium supplemented with different concentrations and combinations of BAP, Kn and Zeatin and NAA, IAA, IBA. The highest percentage (86) of shoot regeneration with maximum shoots per culture (12 ± 0.43) was obtained from nodal explants on MS medium supplemented with 0.5 mg/l BAP and 0.2 mg/l NAA. Addition of 10% coconut water and 75 mg/l urea to the medium enhanced shoot multiplication and elongation, respectively. Rooting of the regenerated shoots was optimum on half-strength MS enriched with 3.0 mg/l IBA. Regenerated plantlets were successfully acclimatized. After acclimation, plantlets were transplanted in the experimental field where the survival rate was 80%.

Multiplikasi Tunas Tanaman Kapas (Gossypium spp.) Varietas Kanesia 15 Menggunakan Kombinasi BAP dan NAA secara In Vitro

Tujuan penelitian ini untuk mengetahui mengenai multiplikasi tunas tanaman kapas (gossypium spp.) varietas Kanesia 15 menggunakan kombinasi BAP dan NAA secara in vitro. Penelitian ini dilaksanakan pada bulan Oktober 2017 Sampai Dengan Januari 2018 bertempat di Laboratorium Kultur Jaringan Politeknik Negeri Jember. Rancangan percobaan yang digunakan adalah Rancangan Acak Lengkap (RAL) Faktorial yang terdiri atas 2 faktor dengan 4 ulangan. Faktor pertama adalah BAP (B) yaitu B1= 0,00 ppm, B2= 1,50 ppm, B3= 3,00 ppm, dan B4= 4,50 ppm. Faktor kedua adalah zat pengatur tumbuh NAA (N) yaitu N1= 0,00 ppm, N2= 0,10 ppm, dan N3= 0,50 ppm. Data pengamatan yang diperoleh, diuji menggunakan Analysis of Varians (ANOVA), jika berpengaruh nyata dilanjutkan dengan uji lanjut Duncan Multiple Range Test (DMRT) pada taraf 5%. Hasil penelitian menunjukkan bahwa pemberian BAP pada media memberikan pengaruh yang berbeda tidak nyata terhadap kedinian tunas, jumlah tunas, dan diameter eksplan, namun memberikan pengaruh yang sangat nyata terhadap tinggi eksplan, jumlah ruas, jumlah akar, dan panjang akar.

In vitro callus induction and indirect organogenesis of Brucea mollis Wall. ex Kurz – A potential medicinal plant of Northeast India

The present investigation was carried out to develop an efficient protocol for in vitro propagation of Brucea mollis Wall. ex Kurz, an important medicinal plant, confined to only Karbi Anglong district of Assam. The plant is extensively used to cure malaria. Due to several natural factors like inefficient pollination, seed dispersal mechanisms and anthropogenic activities, the plant is facing extinction. Through indirect organogenesis from leaf and internode explants on MS (Murashige and Skoog) media, callus induction was achieved at a maximum (100%). However, the minimum time (7–15 days) was observed for induction of callus on B5 as compared to MS media. The color and texture of the callus varied depending on the type of explants as well as the concentrations of growth regulators used in both the media. Best response for percentage of callus induction (100%) was obtained from the leaf explants on MS media, when the media was supplemented with the four combinations of growth regulators, i.e., (i) BAP (8.88 μM) + NAA (1.61 μM), (ii)BAP (8.88 μM) + NAA (2.68 μM), (iii) BAP (8.88 μM) + 2,4-D (2.27 μM) and (iv) BAP (8.88 μM) + 2,4-D (4.54 μM). While in the case of internode explants, maximum (100%) percentage for callus induction was obtained with BAP (8.88 μM) + NAA (1.61 μM) and BAP (8.88 μM) + NAA (2.68 μM). Multiple shoot regeneration was achieved from the callus of all the three types of explants when transferred to shoot regeneration media (both MS and B5 media) supplemented with BAP, NAA, Kinetin and IBA at different concentrations and combinations. The maximum percentage of shoot regeneration (100%) from the calli of all explant types were obtained with the combination of BAP, NAA and IBA. However, the best response in terms of minimum number of days taken to regenerate shoots and maximum shoot length was obtained using the leaf explants at BAP (8.88 μM) + NAA (2.68 μM) + Kinetin (13.95 μM) on MS media. The regenerated shoots were cultured for rooting on half strength media supplemented with IBA and NAA singly, and the highest rooting percentage was achieved when MS media was supplemented with 14.76 μM IBA. About 60% of the plantlets survived during acclimatization and were successfully transferred to the field. The regeneration protocols standardized for B. mollis could be found most effective for mass propagation of the medicinally important plant as well as its conservation.

Micropropagation of ‘Chester thornless’ Blackberry Cultivar using Axillary Bud Explants

Sexual propagation is not suitable for production of true to type plants for commercial cultivation of selected cultivars due to the having heterozygous characteristics. Rubus species are highly heterozygous and therefore when they are propagated by seeds the high plant variability is obtained. Micropropagation methods became very important and have many advantages compared to the classical propagation methods. In Turkey, blackberries are commonly propagated by hard and soft wood cuttings, by layering and bush division. Propagation by layers requires rather large area for a layered, and weed control among the layers is a problem. However, propagation by cuttings is connected with important problem in inadequate rooting, especially in ‘Chester Thornless’ cultivar, which is one of the most commercially grown cultivar. In addition, limited number of plants can be propagated using classical propagation method and thus demand of growers for the plants can not be satisfied. Optimized and highly efficient regeneration protocols are also important for genetic transformation for improving of new cultivars. In this study we focused on optimization of protocol for in vitro propagation of ‘Chester Thornless’ blackberry cultivar using nodal segments. For this purpose, explants were cultured on Woody Plant Medium (WPM) supplemented with combination of plant growth regulators BAP (1.5, 2, 3 mg/l) with NAA (0, 0.1, 0.2, 0.4 mg/l) to detect the best multiplication medium. According to the obtained results, the highest proliferation rate was obtained for cultivar ‘Chester Thornless’ that produced 9.66 shoots on a medium containing 2 mg/I BAP+0.2 mg/I IBA combination. GA3 effects on multiplication rates were also studied and generally the highest rates obtained by while GA3 were not used. For the rooting, different concentrations of IBA and NAA (0, 0.1, 0.2 and 0.4 mg/I) were compared. A concentration of 0.4 mg/l NAA gave the greatest number of roots and maximum root length.

Effect of IBA and BAP on Shoot Growth of Tawangmangu Tangerine (Citrus reticulate) by In-VitroEffect of IBA and BAP on Shoot Growth of Tawangmangu Tangerine (Citrus reticulate) by In-Vitro

Tawangmangu tangerine (Citrus reticulate Blanco subsp. tawangmangu) is one of prime local fruit from Tawangmangu region, Karanganyar, Central Java. This tangerine have good appearance, easy to peeled, sweet flavored fruit, and high production rate. However at 1984 Tawangmangu tangerine run into depreciation of population caused by CVPD (Citrus Vein Phloem Degeneration). As a result, this variety listed as endangeredThe effort of consevation by culture tissue technique can order to obtain prime seeds that free from virus or disease. This study was aimed to understand the effect of IBA and BAP in culture medium on growth of apical and lateral shoot of Tawangmangu tangerine. The treatment provided were IBA (0 ppm, 0.5 ppm, and 1 ppm) and BAP (0 ppm, 1 ppm and 2 ppm). The treatment in this research was using complete randomized factorial design in 9 different treatment. This research resulted that added combination of IBA and BAP affecting growth of Tawangmangu tangerine but the interaction of IBA and BAP did not happen. The highest shoot length occurs on treatment IBA 1 ppm+BAP 2 ppm while when the fastest shoot appears occurs on treatment IBA 0 ppm+ BAP 2 ppm and the highest shoot formed on treatment IBA 0.5 ppm+BAP 2 ppm. Benefit of the research Efforts to conserve Tawangmangu orange plants need to be done to maintain the diversity of germplasm and support the cultivation in a manner. One technique that can support the conservation of plants is by tissue culture techniques.