Genetic homogeneity revealed in micropropagated Bauhinia racemosa Lam. using gene targeted markers CBDP and SCoT.

Two gene targeted markers i.e. CAAT box-derived polymorphism (CBDP) and start codon targeted (SCoT) polymorphism were applied to analyze the genetic stability of in vitro propagated plants of Bauhinia racemosa Lam. multiplied by enhanced axillary shoot proliferation of mature tree derived nodal explant. Nine randomly selected micropropagated plants of 1 year age were subjected to molecular analysis. The isolated genomic DNA samples were subjected to PCR amplification with a total of 61 primers (25 CBDP and 36 SCoT) out of which 39 primers (21 CBDP and 18 SCoT) produced scorable amplicons. A total of 97 and 88 clear, distinct and reproducible amplicons were produced by CBDP and SCoT primers, respectively. The monomorphic banding pattern obtained through all the tested primers corroborated the true to type nature of in vitro propagated plants of B. racemosa.

Exploring genetic variability in Prosopis cineraria using two gene targeted CAAT box-derived polymorphism (CBDP) and start codon targeted (SCoT) polymorphism markers.

Two gene targeted molecular marker systems, CAAT box-derived polymorphism (CBDP) and start codon targeted (SCoT) polymorphism, were used to assess the genetic diversity and relatedness in Prosopis cineraria, a tree of abiotic stress tolerance, agroforestry and ethano-botanical importance. A total of ten wild populations consisting 49 individuals collected from different locations of Indian Thar Desert were examined for the genetic analysis of P. cineraria. Ten CBDP and seven SCoT primers, total 17 primers, generated 204 bands with an average of 12 bands per primer, of which 159 (76.8%) were polymorphic. The average PIC values for both CBDP and SCoT marker were 0.543 and 0.547, respectively. The cumulative data of these two markers were used to analyze different genetic diversity indices and compute pair-wise distances. The population genetic diversity analysis based on cumulative data of CBDP and SCoT markers revealed the high levels of genetic differentiation (GST = 0.341; GST > 0.15 as high), low value of gene flow (Nm = 0.966; Nm > 1 as high) and high fixation index (FST = 0. 415). The highest genetic diversity was observed among NGBAR populations followed by CHR populations, while SIK populations showed lowest genetic diversity. AMOVA revealed the percent molecular variation was higher within the populations (77%) compared to that of among populations (23%). The clustering pattern based on UPGMA and PCoA plot clearly demonstrated the genetic relationship among the genotypes collected from the different regions of Indian Thar Desert.

Aeroponics for adventitious rhizogenesis in evergreen haloxeric tree Tamarix aphylla (L.) Karst.: influence of exogenous auxins and cutting type.

Tamarix aphylla (L.) Karst., a drought resistant halophyte tree, is an agroforestry species which can be used for reclamation of waterlogged saline and marginal lands. Due to very low seed viability and unsuitable conditions for seed germination, the tree is becoming rare in Indian Thar desert. Present study concerns the evaluation of aeroponics technique for vegetative propagation of T. aphylla. Effect of various exogenous auxins (indole-3-acetic acid, indole-3-butyric acid, naphthalene acetic acid) at different concentrations (0.0, 1.0, 2.0, 3.0, 5.0, 10.0 mg l-1) was examined for induction of adventitious rooting and other morphological features. Among all three auxins tested individually, maximum rooting response (79%) was observed with IBA 2.0 mg l-1. However, stem cuttings treated with a combination of auxins (2.0 mg l-1 IBA and 1.0 mg l-1 IAA) for 15 min resulted in 87% of rooting response. Among three types of stem cuttings (apical shoot, newly sprouted cuttings, mature stem cuttings), maximum rooting (~ 90%) was observed on mature stem cuttings. Number of roots and root length were significantly higher in aeroponically rooted stem cuttings as compared to stem cuttings rooted in soil conditions. Successfully rooted and sprouted plants were transferred to polybags with 95% survival rate. This is the first report on aeroponic culture of Tamarix aphylla which can be utilized in agroforestry practices, marginal land reclamation and physiological studies.

In vitro propagation, ex vitro rooting and leaf micromorphology of Bauhinia racemosa Lam.: a leguminous tree with medicinal values.

A micropropagation system for Bauhinia racemosa Lam. was developed involving axillary shoot proliferation and ex vitro rooting using nodal explants obtained from mature tree. MS medium with 3.0 mg l-1 BA (6-benzyladenine) was optimum for shoot bud induction. For shoot multiplication, mother explants were transferred repeatedly on medium containing low concentration of BA (0.75 mg l-1). Number of shoots was increased up to two passages and decreased thereafter. Shoot multiplication was further enhanced on MS medium containing 0.25 mg l-1 each of BA and Kin (Kinetin) with 0.1 mg l-1 of NAA (α-naphthalene acetic acid). Addition of 0.004 mg l-1 TDZ (thidiazuron) increased the rate of shoot multiplication and 21.81 ± 1.26 shoots per culture vessel were obtained. In vitro regenerated shoots were rooted under ex vitro conditions treated with 400 mg l-1 IBA (indole-3-butyric acid) for 7 min on sterile soilrite. After successful hardening in greenhouse, ex vitro rooted plants were transferred to the field conditions with ≈85% of survival rate. Micromorphological changes were observed on leaf surface i.e. development of vein density and trichomes and stomatal appearance, when plants were subjected to environmental conditions. This is the first report on in vitro regeneration of B. racemosa from mature tree.

In vitro propagation of female Ephedra foliata Boiss. & Kotschy ex Boiss.: an endemic and threatened Gymnosperm of the Thar Desert.

Ephedra foliata Boiss. & Kotschy ex Boiss., (family - Ephedraceae), is an ecologically and economically important threatened Gymnosperm of the Indian Thar Desert. A method for micropropagation of E. foliata using nodal explant of mature female plant has been developed. Maximum bud-break (90 %) of the explant was obtained on MS medium supplemented with 1.5 mg l(-1) of benzyl adenine (BA) + additives. Explant produces 5.3 ± 0.40 shoots from single node with 3.25 ± 0.29 cm length. The multiplication of shoots in culture was affected by salt composition of media, types and concentrations of plant growth regulators (PGR's) and their interactions, time of transfer of the cultures. Maximum number of shoots (26.3 ± 0.82 per culture vessel) were regenerated on MS medium modified by reducing the concentration of nitrates to half supplemented with 200 mg l(-1) ammonium sulphate {(NH4) 2SO4} (MMS3) + BA (0.25 mg l(-1)), Kinetin (Kin; 0.25 mg l(-1)), Indole-3-acetic acid (IAA; 0.1 mg l(-1)) and additives. The in vitro produced shoots rooted under ex vitro on soilrite moistened with one-fourth strength of MS macro salts in screw cap bottles by treating the shoot base (s) with 500 mg l(-1) of Indole-3-butyric acid (IBA) for 5 min. The micropropagated plants were hardened in the green house. The described protocol can be applicable for (i) large scale plant production (ii) establishment of plants in natural habitat and (iii) germplasm conservation of this endemic Gymnosperm of arid regions.

Evaluation of aeroponics for clonal propagation of Caralluma edulis, Leptadenia reticulata and Tylophora indica - three threatened medicinal Asclepiads.

The present study explores the potential of aeroponic system for clonal propagation of Caralluma edulis (Paimpa) a rare, threatened and endemic edible species, Leptadenia reticulata (Jeewanti), a threatened liana used as promoter of health and Tylophora indica (Burm.f.) Merill, a valuable medicinal climber. Experiments were conducted to asses the effect of exogenous auxin (naphthalene acetic acid, indole-3-butyric acid, indole-3-acetic acid) and auxin concentrations (0.0, 0.5, 1, 2, 3, 4 or 5gl(-1)) on various root morphological traits of cuttings in the aeroponic chamber. Amongst all the auxins tested, significant effects on the length, number and percentage of rooting was observed in IBA treated nodal cuttings. Cent per cent of the stem cuttings of C. edulis rooted if pre-treated with 2.0 gl(-1) of IBA for 5 min while 97.7 % of the stem cuttings of L. reticulata and 93.33 % of stem cuttings of Tylophora indica rooted with pre-treatment of 3.0 gl(-1) of IBA for 5 min. Presence of at least two leaves on the nodal cuttings of L. reticulata and T. indica was found to be a prerequisite for root induction. In all the species, the number of adventitious roots per cutting and the percentage of cuttings rooted aeroponically were significantly higher than the soil grown stem cuttings. Shoot growth measured in terms of shoot length was significantly higher in cuttings rooted aeroponically as compared to the cuttings rooted under soil conditions. All the plants sprouted and rooted aeroponically survived on transfer to soil. This is the first report of clonal propagation in an aeroponic system for these plants. This study suggests aeroponics as an economic method for rapid root induction and clonal propagation of these three endangered and medicinally important plants which require focused efforts on conservation and sustainable utilization.