Production of herbicide-resistant cowpea (Vigna unguiculata L.) transformed with the bar gene

Plant genetic engineering has opened new avenues to modify crops and has provided a powerful tool for crop improvement. The present study reports the development of regeneration and genetic transformation protocol for the Turkish cowpea cultivar Akkiz. The immature cotyledons were cultured on Murashige and Skoog (MS) medium containing 0.25–0.75 mg L–1 6-benzylaminopurine with or without 0.25 mg L–1 ?-naphthalene acetic acid. Shoot regeneration varied 44.4%–83.3% with 2.1–5.0 shoots per explant. Regenerated shoots were rooted in MS medium containing 0.50 mg L–1 indole-3-butyric acid and acclimatized in the greenhouse, where they flowered and set seeds. Immature cotyledons were transformed with Agrobacterium tumefaciens strain LBA4404 harboring the recombinant binary vector pRGG containing an herbicide tolerance gene (bar) along with a uida (GUS) gene under 35S promoter. Phosphinothricin was used as a selectable marker at a concentration of 2.5 mg L–1. Putative transformants were screened by the histochemical GUS assay. Furthermore, molecular analysis revealed the presence of the introduced gene in the genome of cowpea cultivar Akkiz. The selected transgenic plants showed a resistance to Basta® nonselective herbicide at up to 10 mL L–1 of water. Putative transgenic plants retained their pigmentation and continued to grow in the greenhouse

Comparison of Shoot Regeneration on Different Concentrations of Thidiazuron from Shoot Tip Explant of Cowpea on Gelrite and Agar Containing Medium

Cowpea (Vigna unguiculata L. Walp) is an important legume grown all over the world as grain crop, animal fodder, cover crop, gren manure and vegetable. The present study compares effects of agar and gelrite on micropropagation from shoot tip explant of two Turkish cowpea cultivars Akkiz and Karagoz using 0.15, 0.15, 0.35 mg/l Thidiazuron (TDZ), 3 g/l activated charcoal, 2 mg/l yeast extract with and without 1.25 mg/l Polyvinylpyrrolidone (PVP). To overcome problem of endogenic bacterial contaminations, all cultures contained 500 mg/l augmentin and incubated at 24 ± 2o C in 16 h light photoperiod for eight weeks. Therafter, all explants were transferred to MS medium for two weeks for shoot regeneration and elongation under same incubation and photoperiod conditions. The results showed that frequency of shoot regeneration increased with increase in TDZ concentrations in both cultivars on both agar and gelrite gelled medium. Both cultivars showed maximum mean number of shoots per explant in gelrite compared to agar gelled medium. Maximum number of 4.72 and 2.86 shoots per explant were recorded on MS medium containing 0.25 mg/l TDZ in cv. Akkiz and cv. Karagoz respectively. Hyperhydricity was recorded on some regenerated shoots, which was more prominant on agar. Agar gelled medium had greater shoot length compared to gelrite medium in both cultivars. Regenerated shoots were rooted easily on MS medium containing 0.50 mg/l IBA with regeneration of mean number of 4 secondary shoots on cv. Akkiz and 3 on cv. Karagoz. Rooted plantlets were successfully hardened in the growth chamber and subsequently established in the greenhouse; where they flowered and set seeds. The recorded survival rate of the plants was 100%. Plants looked healthy with no visible detecTab. phenotypic variations

Factors affecting efficient in vitro micropropagation of Muscari muscarimi Medikus using twin bulb scale

AbstractEndemic Muscari muscarimi Medikus is the most fragrant plant among Muscari species and has a high ornamental potential. The natural populations of M. muscarimi, are severely affected by increased environmental pollution and urbanization. There is a need to develop a micropropagation method that should serve effectively for commercial propagation and conservation. Therefore, the study targeted to set up a strategy for efficient in vitro bulblet regeneration system of M. muscarimi using twin scale bulb explants on 1.0×MS medium containing 4.44, 8.88, 17.76μM BAP (6-Benzylaminopurine) plus 2.685, 5.37, 10.74μM NAA (α-Naphthalene acetic acid). Maximum number of 19 daughter axillary bulblets and 16 daughter adventitious bulblets per twin bulb scale explant was regenerated on 1.0×MS medium containing 17.76μM BAP plus 10.74μM NAA and 17.76μM BAP plus 2.685μM NAA respectively. The daughter bulblets regenerated on twin bulb scales on 8 out of 9 regeneration treatment could be easily rooted on 1.0×MS medium containing 4.9μM IBA (Indole-3-butyric acid). The daughter bulblets regenerated on 9th treatment (1.0×MS medium containing 17.76μM BAP plus 10.74μM NAA) were transferred to 1.0×MS medium containing 30g/l sucrose to break negative carry over effect of this dose of BAP–NAA, where they grew 2–3 roots of variable length. Daughter bulblet diameter was increased by culturing them on 1.0×MS medium containing 4.44μM BAP plus 5.37μM NAA. The results verified that both age and the source of explants had significant effect on regeneration. In another set of experiments, twin scales were obtained from in vitro regenerated daughter bulblets, although they induced bulblets, yet their bulblet regeneration percentage, mean number of bulblets per explant and their diameter were significantly reduced. In vitro regenerated bulblets were acclimatized in growth chamber under ambient conditions of temperature and humidity on peat moss, where they flowered. The study provides important information about selection of suitable micropropagation medium, strategies to improve bulblet diameter and rooting of M. muscarimi which offers a scope for commercial propagation

Optimization of Sterilization and Micropropagation of Water Lettuce (Pistia stratiotes L.)

Water lettuce (Pistia stratiotes) is an important medicinal aquatic plant that is also used for phytoremediation or as bioindicator. It is very difficult to multiply the plant using micropropagation techniques due to high contaminations. The study aimed to alleviate this problem for efficient multiplication of the plant subsequently. The study made use of 60% commercial bleach (5% NaOCl) or 60% H2O2 for surface sterilization for 10 min followed by adding 500 mg/l Duocid® (a wide spectrum antibiotic) in the culture medium. The results showed that commercial bleach was not proper sterliser as all explants bleached; whereas, 60% H2O2 based surface sterilization was most optimum for the purpose. The Surface sterilized plants were in vitro cultured on agar solidified or liquid MS medium containing 0.05-0.40 mg/l kinetin. Cent percent shoot regeneration frequency was on both agar solidified or liquid medium. However, all explants on agar solidified or liquid medium had tenancy to die turning all of them to albino. Therefore, multiplication under ex vitro conditions on liquid MS medium containing 0.05-0.40 mg/l kinetin or BAP using unsterilized shoot meristems without adding sucrose. Subsequently, the explants induced 2.0-40 plantlets per explant on 2.0-3.0 mg/l kinetin or BAP containing culture medium. The plants regenerated on both culture media were comparable with plants growing under natural conditions and did not show any sign of stress causing albinism or necrosis

Current Trends In Fenugreek Bıotechnology And Approaches Towards Its Improvement

Fenugreek (Trigonella foenum-graecum L.) plant is cultivated throughout the world as medicinal, food, condiment, dye, and forage. First biotechnological study was carried out in 1945 that reported the effects of diniconazole, a triazole-type fungicide on cell suspension cultures of fenugreek. It was followed by many studies using cell suspension and callus culture emphasising increased production of protein and economically important metabolites like trigonelline, sapogenin, Isoflavonoid pterocarpans, diosgenin, gitogenin and tigogenin from callus, leaves, stems and roots explants. Plant tissue culture studies have emphasied use of callus, cotyloden, hypocotyls and shoot tip epicotyls, apical meristem, cotyledon node and cotyledon leaf explants. Most of the researchers agree difficulty in in vitro rooting. Protoplast studies have also been reported using leaf mesophyll and root apices. Leaf mesophyl protoplasts could be converted to leafy shoots whereas, root apices protoplasts gave cell colonies or roots only. Genetic transformation studies using Agrobacterium rhizogenes and A. tumefaciens are at initial stages and the genes used in the studies only present transformation with either marker or reporter genes. There is a single report on molecular charachterisation of fenugreek from India using 10 RAPD and ISSR primers that revealed interspecific polymorphysm. A review of all this suggests that plant biotechnology of fenugreek is at initial stages of development and there is lot to do for the improvement and breeding of fenugreek

Genetic transformation of common beans (Phaseolus vulgaris L.) through Agrobacterium tumefaciens carrying Cry1Ab gene

Background: Seed beetles are one of the most important causes of yield loss in bean production. It is essential to develop resistant varieties in the fight against these pests. Agrobacterium-based gene transformation is the most widely used breeding method worldwide to develop insect-resistant varieties. Methods and results: Embryonic axes and plumule explants were obtained from Agrobacterium tumefciens treated mature zygotic embryos of low and high raw protein-based common bean cultivars Akman 98 and Karacaşehir 90. Agrobacterium tumefaciens contained a synthetic Bacillus thuringiensis insecticidal crystal protein gene (Bt Cry1Ab) controlled by the 35S promoter and NOS terminator sequences. The transformation event was genotype and explant dependent. The plumule explants could not withstand kanamycin-based selection pressure and died. It was possible to get two transgenic plants using embryonic axis explants of low protein cultivar Akman 98. These results were validated using GUS analysis, PCR, RT-PCR, bioassay analysis, and ELISA test from the samples taken from T0 and T1 generations. Bioassay tests showed that these plants were protected from the damage of legume seed insects (Bruchus spp.). Conclusions: The results are very encouraging and may help in producing better transgenic common bean germplasm leading to safe agriculture and reducing environmental pollutions. © 2022, The Author(s), under exclusive licence to Springer Nature B.V

Microelement Contents and Fatty Acid Compositions of some Isatis Species Seeds

Isatis species are important biennial, herbaceous shrubs represented by 40 taxa, of which 24 are endemic to Turkey. The chemical compounds found in leaves and roots of all Isatis species have medicinal properties and are used for treatment of a wide range of disorders. No study reports seed characteristics, microelement contents and fatty acid compositions of the seeds of any Isatis species except I. tinctoria. Therefore the study was carried out to find seed fatty oil, fatty acid compositions protein content, characteristics, and microelement contents of I. aucheri, I. cochlearis, I. constricta, I. demiriziana (endemic), I. glauca and I. lusitanica. The results showed fatty oil and protein content of these species in range of 2 to 10% and 16.07 to 25.17%, respectively. Furthermore, these seeds were rich in microelements Al, Ca, Fe, K, Mg, Na and P. Major fatty oil components of these species were determined as erucic, oleic, linoleic and linolenic acid; such that erucic acid content of these species ranged 16.94 to 30.52%

Direct Axillary Shoot Regeneration From The Mature Seed Explant Of The Hairy Vetch (Vicia Villosa Roth)

The hairy vetch (Vicia villosa Roth) is a climbing, prostrate or trailing legume grown as forage.It fixes atmospheric nitrogen, reduces soil erosion and provides an instant mulch. Multiple axillary shoot regeneration from a mature seed explant (zygotic embryo with two cotyledons) was obtained on MS medium containing 0.05 – 1.6 mg/l TDZ with or without 0.10 mg/l IBA. The frequency (%) of shoot regeneration ranged from 45.83-75.00% with a maximum number of 28.6 shoots per explant on MS medium containing 0.20 mg/l TDZ-0.10 mg/l IBA. The mean shoot length decreased proportionately with each increase in TDZ concentration irrespective of the IBA concentration in the culture medium. However, comparing the two types of regeneration media, longer shoots were recorded in the presence of IBA in the culture medium. Regenerated shoots were pulse treated with 50 mg/l IBA for 5, 10 and 20 min for rooting

3-Benzyl­isochroman-1-one

In the mol­ecule of the title compound, C16H14O2, the aromatic rings are oriented at a dihedral angle of 78.49 (3)°. The heterocyclic ring adopts a twist conformation. In the crystal structure, inter­molecular C—H⋯O hydrogen bonds link the mol­ecules into chains along the c axis

Ethnobotanical Uses of Roots of Various Plant Species in Turkey

Turkey has advantage of lying on transection of three climatic zones. Namely Europe-Siberia, Iran- Turan and the Mediterranean region situated between 26 and 45° east longitude and 36–42° north latitude in the Northern hemisphere. The number of plant species subspecies, taxa and varieties in Flora of Turkey is above 12,000. In terms of plant diversity in the temperate zone, it attracts attention with its features that are different from the neighbouring countries around it in Asia, Europe and the Middle East. This has led to the development of many distinct ethno-medicinal-botanical habits among local people; who use different plant parts like roots, leaves, flowers, fruits, herbs, seeds, etc. in their cuisines, natural dyeing, decoration, textile dyeing and medicinal purposes, etc. This study reviews ethnomedicinal and botanic uses of the 196 taxa belonging to 54 families and 113 genera grown in Turkey