Browsing by Author "Uma Jaiswal"

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6-Benzylaminopurine mediated indirect organogenesis in Sapindus trifoliatus L. through internodal segments
(Springer, 2025) Pooja Asthana; Manoj K. Rai; Uma Jaiswal
The effect of plant growth regulators (PGRs), explant orientation and subculturing of callus for different passages on the callus induction and shoot differentiation in a medicinal tree Sapindus trifoliatus L. were investigated. The callus was induced from internodal explants obtained from a mature tree on MS medium containing different PGRs, i.e., 2,4-D, BAP, Kin, and zeatin. However, shoot differentiation from callus was obtained only on a medium containing BAP (0.1–3.0 mg l−1) or higher concentrations of zeatin (3.0 or 5.0 mg l−1). The highest number of shoots was obtained when the callus induced on the medium fortified with 3.0 mg l−1 BAP was transferred to a medium containing a comparatively lower concentration (1.0 mg l−1) of BAP. The orientation of the explant (i.e., horizontal or vertical) did not affect callus initiation and further shoot differentiation significantly. The subculturing of callus for different passages had a pronounced effect on shoot differentiation from the callus. The internodal callus showed 100% response for shoot differentiation and an increase in shoot numbers up to the fourth subculture; but, after that significant decline in response and shoot number. The maximum number of shoots (11.43 ± 0.79) was produced after the subculture up to the fourth passage. Thus, by adopting the approach of harvesting shoots and subculture of the remaining callus for seven passages, an average of 45 shoots per 500 mg callus culture was obtained. Maximum percent rooting in micro shoots was obtained when they were pulse-treated with 0.5 mg l−1 IBA for 24 h and transferred to a full-strength MS medium without auxins. Histological analysis revealed the well-developed apical meristem-like structure with leaf primordia during adventitious shoot bud development. The plants developed through internodal callus were established successfully in soil with a 90% survival rate. © The Author(s) under exclusive licence to Society for Plant Research 2024.
6-Benzylaminopurine mediated indirect organogenesis in Sapindus trifoliatus L. through internodal segments
(Springer, 2024) Pooja Asthana; Manoj K. Rai; Uma Jaiswal
The effect of plant growth regulators (PGRs), explant orientation and subculturing of callus for different passages on the callus induction and shoot differentiation in a medicinal tree Sapindus trifoliatus L. were investigated. The callus was induced from internodal explants obtained from a mature tree on MS medium containing different PGRs, i.e., 2,4-D, BAP, Kin, and zeatin. However, shoot differentiation from callus was obtained only on a medium containing BAP (0.1–3.0 mg l−1) or higher concentrations of zeatin (3.0 or 5.0 mg l−1). The highest number of shoots was obtained when the callus induced on the medium fortified with 3.0 mg l−1 BAP was transferred to a medium containing a comparatively lower concentration (1.0 mg l−1) of BAP. The orientation of the explant (i.e., horizontal or vertical) did not affect callus initiation and further shoot differentiation significantly. The subculturing of callus for different passages had a pronounced effect on shoot differentiation from the callus. The internodal callus showed 100% response for shoot differentiation and an increase in shoot numbers up to the fourth subculture; but, after that significant decline in response and shoot number. The maximum number of shoots (11.43 ± 0.79) was produced after the subculture up to the fourth passage. Thus, by adopting the approach of harvesting shoots and subculture of the remaining callus for seven passages, an average of 45 shoots per 500 mg callus culture was obtained. Maximum percent rooting in micro shoots was obtained when they were pulse-treated with 0.5 mg l−1 IBA for 24 h and transferred to a full-strength MS medium without auxins. Histological analysis revealed the well-developed apical meristem-like structure with leaf primordia during adventitious shoot bud development. The plants developed through internodal callus were established successfully in soil with a 90% survival rate. © The Author(s) under exclusive licence to Society for Plant Research 2024.
Alginate-encapsulation of nodal segments of guava (Psidium guajava L.) for germplasm exchange and distribution
(Headley Brothers Ltd, 2008) Manoj K. Rai; Vijai S. Jaiswal; Uma Jaiswal
Nodal segments obtained from in vitro grown plantlets of guava (Psidium guajava L.) were encapsulated in calcium alginate beads for germplasm exchange and distribution. The best gel complex for encapsulation of nodal segments was achieved using 3% (w/v) sodium alginate and 100 mM calcium chloride. The maximum conversion of encapsulated nodal segments into plantlets was obtained on growth regulator-free, full-strength, liquid Murashige and Skoog medium after a pulse treatment with 4.4 μM BAP (6-benzylaminopurine) for 1 week prior to encapsulation. Plants regenerated from encapsulated nodal segments were acclimatised successfully. The present encapsulation approach may also be useful in large-scale propagation of desirable elite genotypes and genetically modified plants.
An improved method of proliferation of proembryogenic calli of Mangifera indica L. var. Amrapali for scale-up of somatic embryo production
(2004) Hussain Ara; Uma Jaiswal; V.S. Jaiswal
Rapid enhancement of proliferation of PECs (proembryogenic calli), obtained from the excised nucellus tissue of var. Amrapali, with high efficiency somatic embryo production has been achieved by manipulating the medium with different growth regulators. Among the growth regulators tested, 1. 0 mg/l 2,4-D (2,4-dichlorophenoxy acetic acid) and 1.0 mg/l NAA (α-naphthalene acetic acid), either alone or with 1.0 mg/l Kn (kinetin), stimulated proliferation of the PEC in both the liquid and solid medium; although, it was more profuse (<5 times) in the liquid medium. Depending upon the physical state and growth regulator(s) supplemented to the medium, the PECs appeared in three morphotypes. Type-III PEC, which was in the form of fine suspensions of dispersible and highly proliferating single cells and small cell aggregates, showed greater potentially for differentiation of somatic embryos in both states of the medium; however, in liquid medium the globular somatic embryos dedifferentiated and callused soon after their differentiation. In contrast, on the semi-solid medium these globular somatic embryos successfully developed into cotyledonary-stage. Type-III PEC (10 mg) had the capacity to produce up to 350 somatic embryos on semi-solid medium. Such somatic embryos matured and grew into plantlets. The method offers a possibility of rapid multiplication in the mango var. Amrapali.
In vitro regeneration and improvement in tropical fruit trees: An assessment
(Springer Netherlands, 2005) Madhulika Singh; Uma Jaiswal; V.S. Jaiswal
In vitro regeneration protocol has been developed for many tropical fruit trees by using juvenile as well as mature explants. Regeneration via somatic embryogenesis have been obtained in a number of cases e.g., while in citrus, sugar apple and papaya, etc. induction of androgenic haploids are successful, in guava and feijoa only callus results in anther cultures. Somaclones have helped in the selection of seedless Musa. Synthetic seed technology has aided in raising plantlets from encapsulated embryos of guava, mango, papaya, etc. Gene transfer techniques can further prove to be useful in the improvement of varieties. © 2005 Springer Science + Business Media, Inc. All rights reserved.
In vitro selection of NaCl-tolerant callus lines and regeneration of plantlets in a bamboo (Dendrocalamus strictus Nees.)
(2003) Madhulika Singh; Uma Jaiswal; Vijai Shanker Jaiswal
Sodium chloride-tolerant plantlets of Dendrocalamus strictus were regenerated successfully from NaCl-tolerant embryogenic callus via somatic embryogenesis. The selection of embryogenic callus tolerant to 100 mM NaCl was made by exposing the callus to increasing (0-200 mM) concentrations of NaCl in Murashige and Skoog medium having 3% (w/v) sucrose, 0.8% (w/v) agar, 3.0 mg1-1 (13.6 μM) 2,4-dichlorophenoxyacetic acid (2,4-D), and 0.5mgl-1 (2.3 μM) kinetin (callus initiation medium). The tolerance of the selected embryogenic callus to 100 mM NaCl was stable through three successive transfers on NaCl-free callus initiation medium. The tolerant embryogenic callus had high levels of Na+, sugar, free amino acids, and proline but a slight decline was recorded in K+ level. The stable 100 mM NaCl-tolerant embryogenic callus differentiated somatic embryos on maintenance medium [MS medium + 3% sucrose + 0.8% agar + 2.0 mgl-1 (9.0 μM) 2,4-D + 0.5 mg 1-1 (2.3 μM) kinetin] supplemented with different (0-200 mM) concentrations of NaCl. About 39% of mature somatic embryos tolerant to 100 mM NaCl germinated and converted into plantlets in germination medium [half-strength MS + 2% sucrose + 0.02 mg 1-1 (0.1 μM) α-naphthaleneacetic acid + 0.1 mg 1-1 (0.49 μM) indole-3-butyric acid] containing 100 mM NaCl. Of these plantlets about 31% established well on transplantation into a garden soil and sand (1:1) mixture containing 0.2% (w/w) NaCl.
In vitro selection, regeneration and characterization of NaCl-tolerant plants of Sapindus trifoliatus: an important multipurpose tree
(Springer Science and Business Media B.V., 2023) Pooja Asthana; Manoj K. Rai; Uma Jaiswal
The present study demonstrates the selection of NaCl tolerant callus line and regeneration of NaCl tolerant plants of Sapindus trifoliatus L. from selected callus via somatic embryogenesis. Callus induced from leaf disc explants was subjected to the selection of NaCl-tolerant callus line on MS medium containing NaCl. The exposure of embryogenic callus to NaCl (0–250 mM) allowed the selection of callus to tolerate 100 mM NaCl, and it confirms with stability test by the three consecutive transfers of 100 mM NaCl tolerant callus to a medium devoid of NaCl and re-transfer to 100 mM NaCl supplemented medium. NaCl selected and non-selected embryogenic callus lines were characterized by the analysis of callus growth, tolerance index, and accumulation of both inorganic (Na+ and K+) and organic (proline, glycine betaine and soluble sugars) content. 100 mM NaCl-tolerant callus showed the formation of globular structure, which differentiated different stages of somatic embryos on a maintenance medium containing NaCl (0-200 mM). Mature cotyledonary stage somatic embryos developed from 100 mM NaCl tolerant globular structure were germinated on germination medium containing NaCl. About 67 and 38% NaCl tolerant somatic embryos were germinated on a medium without NaCl and containing 100 mM NaCl, respectively. NaCl tolerant plants developed on different germination medium with NaCl survived when transplanted to 0.2 or 0.3% (w/w) NaCl mixed potting mixture. Six-month-old established 100 mM NaCl tolerant plants were characterized by analyzing various physiological and biochemical parameters i.e., chlorophyll, carotenoid, L-proline content and antioxidative (MDA, SOD, CAT and APx) activities. The results suggest that in vitro selection technique offers an efficient way to develop NaCl tolerant plants of S. trifoliatus. © 2023, The Author(s), under exclusive licence to Springer Nature B.V.
Induction of somatic embryogenesis and plant regeneration from leaf callus of Terminalia arjuna Bedd.
(1998) Nishi Kumari; Uma Jaiswal; V.S. Jaiswal
A protocol for effective plant regeneration via somatic embryogenesis has been developed for Terminalia arjuna Bedd. Calluses were initiated from leaves of mature trees on Murashige and Skoog's medium (MS) supplemented with 5 mg l-1 2,4-dichlorophenoxyacetic acid, 0.01 mg l-1 kinetin, 3% sucrose and 0.8% agar. The calli showed differentiation of globular structures when transferred to the MS basal medium. Globular structures enlarged and produced secondary globular structures and/or somatic embryos. Continued production of globular structures, their differentiation into embryos and germination of embryos occurred on the MS medium with 3% sucrose and 0.8% agar. The plantlets were hardened and transferred to the soil. Such in vitro raised plants showed luxuriant growth in field condition.
Micropropagation of Sapindus trifoliatus L. and assessment of genetic fidelity of micropropagated plants using RAPD analysis
(2011) Pooja Asthana; V.S. Jaiswal; Uma Jaiswal
An efficient in vitro propagation system has been developed for rapid micropropagation of Soapnut (Sapindus trifoliatus Linn.), a medicinally and economically important tree from nodal (axillary bud) segments of seedlings. The frequency of shoot regeneration from seedling node explant was influenced by the age of the seedlings, growth regulators and successive transfer of the mother explant. Explants from 4-week-old seedlings yielded the maximum shoot regeneration frequency (97. 22%) on full-strength MS medium supplemented with 1. 0 mg l-1 of 6-benzylaminopurine (BAP). After harvesting the newly formed shoots, the mother explants transferred to same medium subsequently produced a maximum of 5. 16 shoots per explant after third passage. Further improvement in the morphogenic response occurred when the nodal explants excised from in vitro regenerated shoots were employed, and 6. 89 shoots per explant were obtained on the same medium after the third subculture. Optimal rooting (91. 67%) was obtained by placing the micro-shoots in liquid MS medium with 1. 0 mg l-1 IBA for 24 h and then transferring to the agar solidified MS medium devoid of IBA. The micropropagated shoots with well-developed roots were acclimatized and successfully transplanted to soil with 90% survival rate. Genetic stability of the regenerated plants was assessed using random amplified polymorphic DNA (RAPD). The amplification products were monomorphic in micropropagated plants and similar to those of mother plant. No polymorphism was detected revealing the genetic integrity of micropropagated plants. This is the first report of an efficient protocol for regeneration of S. trifoliatus through organogenesis, which can be applied for further genetic transformation assays and pharmaceutical purposes. © 2011 Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków.
Rooting of microshoots of Mangifera indica L. cv. Amrapali
(1998) Hussain Ara; Uma Jaiswal; V.S. Jaiswal
A two-step protocol has been developed for in vitro rooting of microshoots excised from plantlets obtained from nucellar somatic embryos of Amrapali cultivar of mango. Role of auxins, their concentrations, treatment periods and culture illumination conditions for conferring competence for rooting have been ascertained. Indole-3-butyric acid (IBA) was found to be the most potent among three auxins (IBA, indoleacetic acid and α-naphthaleneacetic acid) tested for gaining competence for efficient rooting and root growth. Maximum rooting (89.71%) was found on auxin-free agar-gelled rooting medium after 24 h pulse treatment with 5.0 mg/l IBA in liquid root induction medium in dark. Dark condition favoured root induction and root growth, whereas light incubation (16 h photoperiod with 60 μE m-2s-1 light intensity) was inhibitory.
Salt tolerant callus lines of Dendrocalamus strictus nees under salt stress: Growth and ion accumulation
(Plant Archives, 2019) Madhulika Singh; V.S. Jaiswal; Uma Jaiswal
Plant tissue culture technique offers a suitable opportunity for development of salt tolerant plants. The technique is also helpful in understanding physiological and molecular mechanisms of tolerance to salt stress under In vitro condition at the cellular, organ, and whole-plant level. This study was performed to understand the growth responses and accumulation of Na+ and K+ ions against the salt stress in NaCl tolerant callus of Dendrocalamus strictus Nees. The embryogenic callus tolerant to 100mM NaCl was screened out by exposing to increasing (0-200mM) concentrations of NaCl in 0.8% (w/v) agar gelled Murashige and Skoog medium having 3% (w/v) sucrose, 13.6 µM 2,4-Dichlorophenoxy acetic acid and 2.3µM Kinetin (Callus initiation medium). Stable growth of NaCl tolerant callus lines on NaCl-free as well as on NaCl-containing medium was recorded. The relative growth rate of NaCl sensitive callus decreased significantly with increasing concentrations of NaCl into the medium while, for tolerant callus was consistent on medium having 50 and 100mM NaCl and decreased further with increasing concentration of NaCl. The FW/DW of NaCl sensitive callus was higher than the NaCl- tolerant callus. The ratio K+/Na+ was recorded low when the callus was grown on medium supplemented with NaCl and among sensitive and tolerant callus this ratio was found least in tolerant callus when they were grown on medium having 100mM NaCl. © 2019 Plant Archives. All rights reserved.
Somatic embryogenesis and plant regeneration from suspension cultures of Sapindus trifoliatus
(Springer Science and Business Media B.V., 2024) Madhulika Singh; Pooja Asthana; Manoj K. Rai; Uma Jaiswal
A procedure for plant regeneration from cell suspension cultures through somatic embryogenesis is described for Sapindus trifoliatus, a commercially and medicinally important tree. Callus was induced from leaf disc on agar-solidified MS medium with 5.0 mg l−1 2, 4-D and 0.01 mg l−1 Kin. Embryogenic cell suspension cultures were established by placing leaf-derived friable calli in PGR-free full-strength MS liquid medium with 3% sucrose. The growth of cell suspension culture was significantly affected by the strength of the MS mineral solution and L-glutamine. Plating of the suspension on semisolid MS medium resulted in the formation of globular structures. These embryogenic globular structures differentiated into secondary globular structures or somatic embryos on a semisolid MS medium. The differentiation of globular structures and different stages of somatic embryos (from globular to cotyledonary) was enhanced by the addition of 200 mg l−1 L-glutamine in the medium. Sucrose at relatively high concentrations (5%) or ABA (0.01 mg l−1) promoted somatic embryo maturation. The highest percentage (about 90%) of germination of somatic embryo and plantlet conversion was achieved on a half-strength MS medium containing 2% sucrose. The plants were hardened and established in soil with a 90% survival rate © The Author(s), under exclusive licence to Springer Nature B.V. 2024.
Somatic embryogenesis and plantlet regeneration in Amrapali and Chausa cultivars of mango (Mangifera indica L.)
(2000) Hussain Ara; Uma Jaiswal; V.S. Jaiswal
Somatic embryogenesis has been obtained from nucellus of two monoembryonic Indian mango cultivars 'Amrapali' and 'Chausa'. Among the four auxins (IAA, IBA, NAA & 2,4-D) tested, only 2,4-D stimulated callus initiation and induction of proembryogenic callus in cultured bisected ovules containing nucellus minus zygotic embryos. The proembryogenic calli produced up to 130 somatic embryos when transferred to 2,4-D-free medium. The presence of 2,4-D in the medium inhibited progression of development of somatic embryos. The best medium for the production, development and maturation of somatic embryos was the modified M4E medium which contained full-strength BS macrosalts, MS microsalts, MS iron-EDTA and MS organics along with 400 mg/1 L-glutamine, 6% (w/v) sucrose and 0.8% (w/v) agar. The mature somatic embryos gave rise to plantlets in liquid medium containing half-strength BS macrosalts and 1.0 mg/1 GA3. The in vitro raised plantlets of Amrapali cultivar have been successfully transplanted in earthen pots containing garden soil, but those of Chausa failed to survive in the garden soil but have been established in pots containing sand and soil (3:1) mixture.
Somatic embryogenesis from sepal explants in Sapindus trifoliatus, a plant valuable in herbal soap industry
(Elsevier B.V., 2017) Pooja Asthana; Manoj K. Rai; Uma Jaiswal
Somatic embryogenesis and complete plant regeneration were achieved in callus cultures derived from sepal explants of Sapindus trifoliatus, an important and valuable tree used in saponin and herbal soap industry. Embryogenic calli were induced on MS medium containing 5.0 mg l−1 2, 4-D and 0.1 mg l−1 Kin. Explant orientation markedly influenced on somatic embryogenesis. The calli induced on medium supplemented with 2, 4-D and Kin showed the formation of whitish or greenish nodular embryogenic structure on MS basal medium containing L-glutamine. These nodular embryogenic structures gave rise to somatic embryos or secondary nodular embryogenic structures or both. Nodular embryogenic structures separated from embryogenic calli and transferred to MS medium containing 200 mg l−1 L-glutamine produced maximum number of globular, heart and cotyledonary stage somatic embryos. Histological studies have revealed the development of different stages of somatic embryos from nodular embryogenic structure. The embryogenic competence of the culture could be maintained by recurrent production of nodular embryogenic structures and somatic embryos. Somatic embryos were germinated and converted into plantlet on MS basal medium containing 2% sucrose. Plants propagated in vitro from somatic embryos were hardened and successfully established in field condition. The improved and promising regeneration method reported here for the S. trifoliatus may be valuable in herbal soap industry. © 2017 Elsevier B.V.
Synthetic seed: Prospects and limitations
(2000) Hussain Ara; Uma Jaiswal; V.S. Jaiswal
The synthetic seed technology has been developed to use somatic embryos and/or other micropropagules as seed analogues successfully in the field or greenhouse, and their mechanical planting at a commercial level. The technology provides methods for preparation of seed analogues called synthetic seeds or artificial seeds from the micropropagules like somatic embryos, axillary shoot buds, apical shoot tips, embryogenic calli as well as protocorm or protocorm-like bodies. For the last fifteen years, intensive research efforts have been made on synthetic seed production in a number of plant species. Despite these researches, practical implementation of the technology is yet to be fully realized due to limitations encountered with the production, development, maturation and subsequent conversion of the micropropagules into plantlets under in vitro or ex vitro conditions. The present article focuses on the technology developed, its achievements and prospects as well as limitations resisting the application of the synthetic seed technology.
Thidiazuron-induced in vitro flowering in Dendrocalamus strictus Nees
(2000) Madhulika Singh; Uma Jaiswal; V.S. Jaiswal
[No abstract available]