In vitro high frequency regeneration through apical shoot proliferation of Hemianthus callitrichoides 'Cuba' - a multipurpose ornamental aquatic plant

WOS:000356358200017Hemianthus callitrichoides 'Cuba' is an ornamental aquatic plant used in water gardens and phytoremediation. A protocol for its rapid and reproducible shoot organogenesis from apical shoot explants was developed. Optimum culture conditions for shoot proliferation were tested in MS media containing different concentrations of 6-(gamma,gamma-dimethylallylamino)purine (2iP), 6-benzylaminopurine (BAP), 1-phenyl-3-1,2,3-thiadiazol-5-yl urea (TDZ), and gibberellic acid (GA(3)). Plant growth regulators significantly (P <= 0.001) affected the diameter, area, and fresh weight of shoot clumps. On the basis of plant growth, the maximum shoot diameter (3.06 cm), area (6.12 cm), and clump weight (2.85 g) of apical shoots were achieved on MS medium containing 0.50 mg L-1 BAP. Higher concentrations of sucrose (9%) showed visible phenotypic growth inhibition and abnormalities were observed in vitro. The regenerated shoots rooted efficiently on MS medium without growth regulators. The growth pattern of multiple shoots indicated their origin from an enlarged shoot base via the proliferation of apical shoots. All in vitro regenerated plantlets acclimatized in aquariums successfully. The simplicity of the protocol and the direct production of multiple shoots make it a potential system that is highly amenable to true-to-type plant regeneration with the ability to maintain genetic stability

Vigna unguiculata bitkisinde dilimlenmemiş ve dikey dilimlenmiş kotiledon boğum eksplantların sürgün rejenerasyonuna etkileri

WOS:000303080400007Vigna unguiculata is an important food legume crop in the semiarid tropics. It suffers from a host of agricultural constraints including damage due to diseases and pests. Judicious application of biotechnological methods can lead to considerable improvement in this important crop. Shoot regeneration from unsliced and longitudinally sliced cotyledon node explants obtained from 3-day-old to 5-day-old seedlings grown in vitro was achieved on Murashige and Skoog (MS) medium containing 1.11, 2.22, 3.33, and 4.44 ?M benzylaminopurine (BA) supplemented with 1.0 mg/L polyvinylpyrrolidone and 500 mg/L bacteriostatic Augmentin. Callus induction was recorded in all culture media on both explants. Th e shoot regeneration frequency (%) of longitudinally sliced cotyledon node explants was 2-fold to 3-fold higher than that of unsliced cotyledon node explants. A maximum number of 9.92 shoots per longitudinally sliced cotyledon node explant was recorded on MS medium containing 3.33 ?M BA. Each increase in the BA concentration of the culture medium proportionally decreased the mean shoot length on both explants. Maximum mean shoot lengths of 2.80 cm on unsliced cotyledon nodes and 3.04 cm on longitudinally sliced cotyledon node explant were recorded on MS medium containing 1.11 ?M BA. Regenerated shoots were rooted on MS rooting medium containing 2.45 ?M indole-3-butyric acid. In vitro regenerated plants were acclimatised at room temperature in growth rooms, where they produced viable seeds.Börülce yarı kurak tropik bölgelerin önemli yemeklik tane baklagil bitkisi olup, bazı zararlara maruz kalmaktadır. Bu bitkinin ıslahı için biyoteknolojik yöntemlerin geliştirilmesi son derecede önemlidir. Bu çalışmada, in vitro koşullarda 3-5 günlük çimlenmiş fi deciklerden elde edilen dilimlenmemiş ve dikey dilimlenmiş kotiledon boğum eksplantları 1,11, 2,22, 3,33 ve 4,44 μM benzylaminopurine (BA), 1,0 mg/L PVP ve 500 mg/L Augmentin içeren MS ortamda kültüre alınarak sürgün rejenerasyonu elde edilmiştir. Tüm ortamlarda her iki eksplant üzerinde de kallus oluşumu gözlenmiştir. Dikey dilimlenmiş kotiledon boğum eksplantlarında dilimlenmemiş kotiledon boğum eksplantlarına göre daha fazla sürgün oluşum görülmüştür. Dilimlenmiş kotiledon boğumunda, en fazla sürgün (9,92 adet) 3,33 μM BA içeren MS ortamından elde edilmiştir. Tüm kültür ortamlarında, BA dozların artışı oransal olarak sürgün uzunluğunun azalmasının sebebi olmuştur. En uzun sürgün oluşumu (2,80 cm ve 3,04 cm), dilimlenmiş ve dilimlenmemiş kotiledon boğumundan 1,11 μM BA içeren MS ortamında görülmüştür. Elde edilen sürgünler 2,45 μM IBA içeren MS ortamda köklendirilmiştir. In vitro koşullarda gelişen bitkilerin adaptasyonu oda sıcaklığında sağlanmış olup, tohum elde edilmişti

Preconditioning effect of cytokinins on in vitro multiplication of embryonic node of grass pea (Lathyrus sativus L.) cultivar Gürbüz

Lathyrus sativus L. (grass pea) is an important grain legume crop grown all over the world. It is very recalcitrant and difficult to regenerate under in vitro conditions. The present study determined the effect of preconditioning of cytokinins [10 and 20 mg L–1 thidiazuron (TDZ) and 2-isopentenyl adenine (2iP)] and postconditioning of dissimilar levels of TDZ-indole-3-butyric acid (IBA) and 2iP-IBA on in vitro shoot multiplication using embryonic node explants. The results showed that preconditioning followed by postconditioning on Murashige and Skoog (MS) medium with 0.25, 0.50, 0.75, and 1.0 mg L–1 2iP or TDZ combined with 0.10 mg L–1 IBA had positive effects on shoot multiplication. It seemed as if the TDZ-preconditioned and TDZ-IBA–postconditioned explants were more regenerative compared to 2iP-preconditioned or 2iP-IBA–postconditioned explants. Maximum shoot induction (100%) was achieved on MS medium containing 1 mg L–1 TDZ and 0.10 mg L–1 IBA, with a mean number of 31.10 shoots per explant. In comparison to the other cytokinin tested, 2iP-preconditioned and 2iP-IBA–postconditioned explants had longer shoots, with a mean shoot length of 8.10 cm. Root initiation was observed on all cultures 4 weeks after the transfer of shoots with the best rooting on MS medium containing 2 mg L–1 IBA. The rooting response was 76.56% with a mean number of 8.03 roots per shoot and root length of 8.2 cm. The healthy plants were transferred to a greenhouse for acclimatization and exhibited 90% survival. It was concluded that TDZ preconditioning was necessary for grass pea multiplication along with TDZ-IBA postconditioning. This regeneration protocol may facilitate future genetic transformation and breeding studies to reduce ß-N-oxalyl-L-?,ß-di-amino propionic acid content in grass pea

Effects of squirting cucumber (Ecballium elaterium) fruit juice on Agrobacterium tumefaciens-mediated transformation of plants

Different concentrations of squirting cucumber (Ecballium elaterium (L.) A.Rich.) fruit juice were added to Agrobacterium tumefaciens growth, leaf disc inoculation, and cocultivation media, to investigate its effect on the transformation frequency of tobacco and potato. A. tumefaciens strain GV2260 harboring p35S GUS-INT and pAoPR1-GUS-INT plasmids were used separately in the transformation experiments. Neomycin phosphotransferase (NPT-II) gene was used as a plant selectable marker at a concentration of 100 mg L–1. The addition of 5–10 mg L–1 squirting cucumber fruit juice to bacterial nutrient medium increased A. tumefaciens growth significantly by 6 h. Moreover, the use of high concentrations (2.5–20 mL L–1) of fruit juice resulted in excessive bacterial growth on cocultivation and selection media around the explants, which was difficult to eliminate by subculture or higher levels of antibiotics. On the other hand, lower concentrations (0.2–1.6 mL L–1) of squirting cucumber fruit juice significantly increased the transformation frequency in both tobacco and potato. Kanamycin-resistant tobacco shoots, rooted in a medium containing 100 mg L–1 kanamycin, were transferred to pots containing organic soil and perlite in growth cabinets for acclimatization. Transgenic plants grew normally and set seeds. The presence of T-DNA in these transformants was confirmed by PCR and GUS analysis