Axillary Branches Research Articles

Transfer of axillary nerve branches to reconstruct elbow extension in tetraplegics: A laboratory investigation of surgical feasibility

In spinal cord injuries at the C6 level, elbow extension is lost and needs reconstruction. Traditionally, elbow extension has been reconstructed by muscle transfers, which improve function only moderately. We have hypothesized that outcomes could be ameliorated by nerve transfers rather than muscle transfers. We anatomically investigated nerve branches to the teres minor and posterior deltoid as donors for transfer to triceps motor branches. In eight formalin-fixed cadavers, the axillary nerve, the teres minor branch, the posterior deltoid branch, the triceps long and upper medial head motor branches, and the thoracodorsal nerve were dissected bilaterally, their diameters measured and their myelinated fibers counted. To simulate surgery, using an axillary approach in two fresh cadavers, we transferred the teres minor or the posterior deltoid branch to the triceps long head and to the thoracodorsal nerve. The posterior division of the axillary nerve gave off the teres minor motor branch and then the branch to the posterior deltoid, terminating as the superior lateral brachial cutaneous nerve. The diameters of the teres minor motor branch, posterior deltoid, triceps long and upper medial head branches, and the thoracodorsal nerve all were ∼2 mm, with minimal variation. The nerves varied little in their numbers of myelinated fibers, being consistently about 1,000. Via an axillary approach, either the teres minor or the posterior deltoid branch could be transferred directly to the thoracodorsal nerve or to triceps branches without any tension.

Bioinformatics Analysis on Regulating Axillary Branch Gene of Cucumber (Cucumis sativus L.)

The regulating axillary branch gene was cloned and named as CsCCD7. Using a series bioinformatic computer softwares, database and online programes, CsCCD7 nucleotide sequence and CsCCD7 amino acid sequence were analyzed and CsCCD7 function was predicted. The results showed that CsCCD7 cDNA full length sequence was 2 136 bp, and included a 1 665 bp ORF which encoded a 554 AA protein; there were 32 kinds of cis-acting regulating element in 2 136 bp cDNA sequence; CsCCD7 was an unstable protein (the unstable coefficient: 40.77), including many phosphorylation sites related with CsCCD7 function; CsCCD7 had no transmembrane domain, and its subcellular localization was in chloroplast. CsCCD7 secondary structure contained four conformations including α-helix, β-sheet, β-turn and random coil. CsCCD7 protein had no signal peptide, so was non-secretory protein and hydrophilicity protein (grand average of hydropathicity: −0.401). Both CsCCD7 secondary and tertiary structure prediction results showed that it was classified as carotenoid oxygenase family. Phylogenetic tree drew by Geneious showed that CsCCD7 was more closely related to AtCCD7 than any other Arabidopsis CCD protein.

Consequences of transforming narrow leafed lupin (Lupinus angustifolius [L.]) with an ipt gene under control of a flower-specific promoter

Phenotypes of five transgenic lines of narrow-leafed lupin (Lupinus angustifolius [L] cv Merrit) stably transformed with the isopentenyl pyrophosphate transferase (ipt) gene from Agrobacterium tumefaciens coupled to a flower-specific promoter (TP12) from Nicotiana tabacum [L.] are described. Expression of the transgene was detected in floral tissues and in shoot apical meristems on all orders of inflorescence. In each transgenic line there was significant axillary bud outgrowth at all nodes on the main stem with pronounced branch development from the more basal nodes in three of the lines. The lowest basal branches developed in a manner similar to the upper stem axillary branches on cv Merrit and bore fruits, which, in two lines, contained a significant yield of filled seeds at maturity. Senescence of the cotyledons was delayed in all lines with green cotyledons persisting beyond anthesis in one case. IPT expression increased cytokinin (CK) levels in flowers, meristem tissues and phloem exudates in a form specific manner, which was suggestive of localized flower and meristem production with significant long-distance re-distribution in phloem. The total number of fruits formed (pod set) on some transgenic lines was increased compared to cv Merrit. Grain size compared to cv Merrit was not significantly altered in transgenic lines.

Partial shoot reiteration in Wollemia nobilis (Araucariaceae) does not arise from ‘axillary meristems’

Conifers are characterized by the paucity of axillary buds which in dicotyledonous trees usually occur at every node. To compensate, conifers also produce 'axillary meristems', which may be stimulated to late development. In juvenile material of Wollemia nobilis (Araucariaceae: Massart's model) first-order (plagiotropic) branches lack both axillary buds and, seemingly, axillary meristems. This contrasts with orthotropic (trunk) axes, which produce branches, either within the terminal bud or as reiterated orthotropic axes originating from axillary meristems. However, plagiotropic axes do produce branches if they are decapitated. This study investigated how this can occur if axillary meristems are not the source. The terminal buds of a series of plagiotropic branches on juvenile trees were decapitated in order to generate axillary shoots. Shoots were culled at about weekly intervals to obtain stages in lateral shoot development. Serial sections were cut with a sliding microtome from the distal end of each sample and scanned sequentially for evidence of axillary meristems and early bud development. Anatomical search produced no clear evidence of pre-existing axillary meristems but did reveal stages of bud initiation. Buds were initiated in a group of small starch-rich cortical cells. Further development involved de-differentiation of these small cells and the development of contrasting outer and inner regions. The outer part becomes meristematic and organizes the apex of the new branch. The inner part develops a callus-like tissue of vacuolated cells within which vascular cambia are developed. This kind of insertion of a branch on the parent axis seems not to have been described before. Axillary meristems in Wollemia characterize the leaf axils of trunk axes so that the origin of reiterated shoots is clear. Plagiotropic axes seemingly lack axillary meristems but still produce axillary branches by distinctive developmental processes. These observations demonstrate limited understanding of branch initiation in trees generally.

Genetic and environmental control of the Verticillium syndrome in Arabidopsis thaliana

BackgroundVerticillium spp. are major pathogens of dicotyledonous plants such as cotton, tomato, olive or oilseed rape. Verticillium symptoms are often ambiguous and influenced by development and environment. The aim of the present study was to define disease and resistance traits of the complex Verticillium longisporum syndrome in Arabidopsis thaliana (L.) Heynh. A genetic approach was used to determine genetic, developmental and environmental factors controlling specific disease and resistance traits and to study their interrelations.ResultsA segregating F2/F3 population originating from ecotypes 'Burren' (Bur) and 'Landsberg erecta' (Ler) was established. Plants were root-dip inoculated and tested under greenhouse conditions. The Verticillium syndrome was dissected into components like systemic spread, stunting, development time and axillary branching. Systemic spread of V. longisporum via colonisation of the shoot was extensive in Ler; Bur showed a high degree of resistance against systemic spread. Fungal colonisation of the shoot apex was determined by (a) determining the percentage of plants from which the fungus could be re-isolated and (b) measuring fungal DNA content with quantitative real-time PCR (qPCR). Four quantitative trait loci (QTL) controlling systemic spread were identified for the percentage of plants showing fungal outgrowth, two of these QTL were confirmed with qPCR data. The degree of colonisation by V. longisporum was negatively correlated with development time. QTL controlling development time showed some overlap with QTL for resistance to systemic spread. Stunting depended on host genotype, development time and seasonal effects. Five QTL controlling this trait were identified which did not co-localize with QTL controlling systemic spread. V. longisporum induced increased axillary branching in Bur; two QTL controlling this reaction were found.ConclusionsSystemic spread of V. longisporum in the host as well as resistance to this major disease trait are described for the first time in natural A. thaliana accessions. This creates the possibility to study a major resistance mechanism against vascular pathogens in this model plant and to clone relevant genes of the involved pathways. Stunting resistance and resistance to systemic spread were controlled by different QTL and should be treated as separate traits. Developmental and environmental effects on pathogenesis and resistance need to be considered when designing and interpreting experiments in research and breeding.

Epigenetics in plant tissue culture

Plants produced vegetatively in tissue culture may differ from the plants from which they have been derived. Two major classes of off-types occur: genetic ones and epigenetic ones. This review is about epigenetic aberrations. We discuss recent studies that have uncovered epigenetic modifications at the molecular level, viz., changes in DNA methylation and alterations of histone methylation or acetylation. Various studies have been carried out with animals, and with plant cells or tissues that have grown in tissue culture but only little work has been done with shoots generated by axillary branching. We present various molecular methods that are being used to measure epigenetic variation. In micropropagated plants mostly differences in DNA methylation have been examined. Epigenetic changes are thought to underlie various well-known tissue-culture phenomena including rejuvenation, habituation, and morphological changes such as flower abnormalities, bushiness, and tumorous outgrowths in, among others, oil palm, gerbera, Zantedeschia and rhododendron.

Anatomical Study of Axillary Artery Variation.

Background: The axillary artery is a direct continuation of the subclavian artery. The axillary artery is usually described as giving off six branches. The first part gives superior thoracic artery. The second part gives lateral thoracic (LT) and thoracoacromial(TAC) arteries. The third part gives three, subscapular(SS), anterior circumflex humeral(ACH)and posterior circumflex humeral(PCH) arteries. Anatomical variations in the branching pattern of axillary artery are quiet common and typically include the subscapular artery(SS), lateral thoracic artery(LT) and the posterior circumflex humeral artery(PCH). The variation of the axillary artery branching pattern has anatomical as well as clinical and surgical relevance given the proximity to the shoulder joint and humerus.Patients & Method: Bilateral axilla dissection was conducted on 26 embalmed axillae (13 cadavers) to allow examination of the axillary artery and its branches. The study was carried out in Department of Human Anatomy, College of Medicine.Results: The classical branching pattern of the LT originating from the axillary artery posterior to the pectoralis minor muscle and the SS producing the circumflex scapular (CS) artery and thoracodorsal(TD) occurred in 77%. The SS was observed originating from the LT 7% of the time. The LT was observed originating from the SS 5% of the time. The LT was observed producing the circumflex scapular artery and TD in the absence of SS 2.5% of the time. The PCH originated from four different sources, from the third part of axillary artery as is classically described in 77%. From the SS 11%. From deep brachial artery DB 9% and from LT 2%.Conclusion: Vascular variation in the axillary artery and its branches is quiet common , This variation should be considered seriously as will implicate risk of bleeding during surgery in the axilla and also the difficulty in interpretation of the angiography after axillary catheterization.

Drought stress mitigation using supplemental irrigation in rainfed chickpea ( Cicer arietinum L.) varieties in Kermanshah, Iran

An experiment was carried out in 2007 to investigate the effects of different irrigation regimes, and chickpea cultivars on chickpea production in the Agricultural Research Station, College of Agriculture, Islamic Azad University, Kermanshah Branch, Iran. The experimental design was split-plot with three replications. Supplemental irrigation at three levels, that is, control treatment (without irrigation) (I0), one time irrigation at 50% flowering stage (I1) and one time irrigation at pod-filling stage(I2), was allocated to main plots and the varieties ILC-482 (V1), Hashem (V2) and Arman (V3) were allotted to sub plots. A significant difference was observed between irrigation treatments in terms of grain yield, plant height, number of axillary branches, distance to the first pod from soil surface, number of grain per plant, number of pod per plant, biological yield, harvest index and 100-grain weight. Such differences were also observed between testing varieties in terms of all traits rather than 100-grain weight. Grain yield was significantly higher for Arman than that of Hashem which was significantly higher than that of ILC-482. Of course, there was no significant difference between Hashem and ILC-482 in terms of grain yield. Arman had the highest values of the number of grain per plant and the highest pod per plant pertained to Arman and Hashem, respectively. High rate of grain yield in irrigation treatment at podfilling stage was associated with yield components, especially with the number of pod per plant and 100-grain weight. Grain yield was positively correlated with number of pod per plant (r = 0.654**), number of grain per plant (r = 0.902**) and 100-grain weight (r = 0.707**). This research showed that podfilling is the most sensitive stage to drought stress, and under water limitation conditions, we can considerably increase grain yield at this stage by one time irrigation, especially for Arman cultivar.

Strigolactones enhance competition between shoot branches by dampening auxin transport

Strigolactones (SLs), or their derivatives, were recently demonstrated to act as endogenous shoot branching inhibitors, but their biosynthesis and mechanism of action are poorly understood. Here we show that the branching phenotype of mutants in the Arabidopsis P450 family member, MAX1, can be fully rescued by strigolactone addition, suggesting that MAX1 acts in SL synthesis. We demonstrate that SLs modulate polar auxin transport to control branching and that both the synthetic SL GR24 and endogenous SL synthesis significantly reduce the basipetal transport of a second branch-regulating hormone, auxin. Importantly, GR24 inhibits branching only in the presence of auxin in the main stem, and enhances competition between two branches on a common stem. Together, these results support two current hypotheses: that auxin moving down the main stem inhibits branch activity by preventing the establishment of auxin transport out of axillary branches; and that SLs act by dampening auxin transport, thus enhancing competition between branches.

Relationship of phenotypic variation in sorghum to nutritive value of crop residues

Improvements in dry matter digestibility of crop residues contribute to greater productivity of livestock consuming dual-purpose sorghums ( Sorghum bicolor L. Moench). Delayed leaf senescence (stay-green), brown-midrib (bmr), leaf disease resistance, and high yield are among sorghum phenotypic traits that could enhance nutritive value of crop residues. The objective was to evaluate nutritive value and hydrocyanic acid potential (HCN-p) of leaf or stem components of crop residues of sorghum lines and hybrids representing differences in phenotypic traits. Four replications of eight or more lines and hybrids made up a randomized complete block design during 2 years in central Texas. Dry matter yield, dry weight loss after extraction in water and incubation in rumen, and fiber concentrations of leaves and stems were measured after grain harvest under field conditions. In addition, HCN-p of axillary branches and mature leaves was quantified. Sample weight loss in water extractions without incubation in rumen was negatively correlated with fiber concentrations, which were measured as the sample weights remaining after extraction in neutral (NDF) and acidic (ADF) detergent solutions. Greater concentrations of water-extractable dry matter and lower concentrations of NDF and ADF were observed for stay-green than for bmr or lines that senesced under drought. Yet, the sum of water-extractable dry matter plus ensuing weight loss during 7 days in rumen, expressed as yield ha −1 of leaf and stem digestible dry matter, was not consistently greater for stay-green than drought-susceptible lines. In addition, the yield of the sum of water-extractable and 7-day rumen losses of stay-green lines was comparable to or less than the bmr line. Under conditions in which leaf senescence varied little, analyses of nutritive value indicated water-extractable dry matter was enhanced and fiber concentrations were reduced in crop residues of stay-green compared to senescent lines.

MOLECULAR ANALYSIS OF MICROPROPAGATED STRAWBERRY PLANTS USING ISSR (INTER SIMPLE SEQUENCE REPEATS) MARKERS FOR ASCERTAINING CLONAL FIDELITY

The present study is aimed at ascertaining the genetic uniformity of the nuclear genome of mother plant and its tissue cultured progenies of strawberry in order to establish true-to-type character through enhanced axillary branching micropropagation by employing ISSR-PCR molecular marker. The study yielded monomorphic bands in all the seven UBC dinucleotide motif primers. The results concluded that the tissue culture raised plantlets of strawberry are genetically identical and clonally uniform at least up to the 15 th culture passage.

Development of a Highly Efficient Micropropagation Method for the Citrus Rootstock ‘Swingle’ Citrumelo [Poncirus trifoliata (L.) Raf. × C. paradisi McFaden

This article describes a complete micropropagation protocol employing node segments from branches of adult trees of ‘Swingle’ citrumelo. Multiple shoots were induced in vitro through enhanced axillary branching. A multiplication rate of 3.79-fold every four weeks was achieved on Murashige and Skoog's media (1962) supplemented with cytokinins (6-benzyl amino purine and kinetin) with 4% sucrose and 0.4% agargel. The highest percent rooting was observed with a combination of indole-3-acetic acid and indole-3-butyric acid. The hardening survival rate of the rooted plantlets was 100% in a soil and agropeat combination. The micropropagated plantlets were tested for their genetic fidelity using seven inter simple sequence repeats primers.

Branching patterns of the lateral thoracic, subscapular, and posterior circumflex humeral arteries and their relationship to the posterior cord of the brachial plexus

Anatomical variations in the branching pattern of the axillary artery are common and typically include the lateral thoracic, subscapular, and the posterior circumflex humeral (PCHA) arteries. Previous investigations of single specimen dissections demonstrate numerous variations to axillary artery branching, but the frequency of these occurrences is unclear. This study quantifies the frequency of variant branching of the lateral thoracic, subscapular, and posterior circumflex humeral arteries, how it they relate to the posterior cord of the brachial plexus. Axillae of 83 cadavers were dissected to allow examination of the axillary artery and its branches. Data were collected observing the branching pattern of the lateral thoracic, subscapular, and posterior circumflex humeral arteries, as well as those branches spatial relationship to the two terminal branches of the posterior cord of the brachial plexus. Some of the more common variations included the thoracodorsal artery arising from the lateral thoracic artery (LTA) (7.2%) and the subscapular artery (SSA) arising from the LTA (5.4%). The SSA also produced the LTA (4.2%) and the PCHA (12%). The PCHA also originated from the deep brachial artery (8.4%) and traversed the triangular interval to supply the deltoid muscle. These findings are relevant to both the anatomical and clinical fields as provide evidence as to the frequency of variant axillary artery branching and the potential for neurovascular elements to exist in a location other than their classical anatomical description.

Analysis of Genetic Stability through Intersimple Sequence Repeats Molecular Markers in Micropropagated Plantlets of Anoectochilus formosanus HAYATA, a Medicinal Plant

Anoectochilus formosanus HAYATA, commonly known as "Jewel Orchids," which has been used as Chinese folk medicines, is being subjected to a huge crisis, whose wild resources have gradually become more and more scarce. Hence, micropropagation protocol by axillary branching established for A. formosanus was employed for large-scale commercial production. In this study, due to the somaclonal variation and on the basis of some virtue of intersimple sequence repeats (ISSR) analysis, we firstly utilized the ISSR primers to investigate the genetic stability of A. formosanus propagated in vitro for a period of more than 5 years. Among the total 100 bands amplified by 17 ISSR markers, 77 bands were distributed in size from 500 bp to 1.5 kbp, while only 5 bands were beyond 1.8 kbp in size. Meanwhile, according to the cluster analysis, genetic similarity was more than 94% and the polymorphism rate was only 2.76% among the total 1810 scorable bands. All results demonstrate A. formosanus, multiplied by axillary branching, maintained high genetic fidelity even after a period of more than 5 years under in vitro propagation with only a low risk of genetic instability. The results from this study provide an important basis for giving evidence of genetic stability of A. formosanus before micropropagation for large-scale commercial production.

The role of AUX1 gene and auxin content to the branching phenotype of Kenaf (Hibiscus cannabinus L.).

The objectives of this research were to identify auxin gene, AUX1, and to determine the plant auxin content and their role in conferring branching on Kenaf. PCR analysis using AUX1 primer capable to amplify the DNA of non branching (KR11) and branching kenaf mutant, resulting in 800 bp PCR product. The sequence of the PCR product showed high degree of homology with the sequence of AUX1 gene of other plants in the NCBI GenBank database, confirming kenaf possession of the gene AUX1. However, some variation on the DNA sequence was found between branching and non branching phenotype indicated allele differences of the same gene which were responsible for the variation in the type of branching. Identification of auxin content in the roots, apical shoot, and axillary branches using spectrophotometry method showed that the branching plant has higher auxin content in the apical shoot compared to the content in the branches. This indicate that AUX1 controls the formation of branches by controlling either the content of auxin in the apical shoot and branches, or the ratio of auxin content in the shoot and branches.

Characterization and Mapping of Rice Mutants, <em>des2</em> and <em>des5</em>

Identification of key regulator genes in the control of spikelet development in panicle, one of the main agronomic traits to determine rice yield, is important to investigate the underlying molecular mechanism and provide the molecular basis for analysis of the agronomic traits. In this study, we have identified a group of mutants with conspicuous reduction of spikelet number, which are designated decreased spikelets ( des ). Two of the des mutants, des2 and des5 , were analyzed in detail. We showed that a single recessive locus was responsible for des2 phenotype, which is located on the long arm of chromosome 6. The des2 locus was cloned and it was found that des2 is a new allele of moc1 . Positional cloning and sequence analysis showed that there is a point mutation in the HLH domain-encoding region of LAX gene in des5 , indicating that des5 is a new allele of lax . Our data indicate that spikelet development is regulated by the same pathway in the control of development of axillary branches in rice.

SlCCD7 controls strigolactone biosynthesis, shoot branching and mycorrhiza-induced apocarotenoid formation in tomato

The regulation of shoot branching is an essential determinant of plant architecture, integrating multiple external and internal signals. One of the signaling pathways regulating branching involves the MAX (more axillary branches) genes. Two of the genes within this pathway, MAX3/CCD7 and MAX4/CCD8, encode carotenoid cleavage enzymes involved in generating a branch-inhibiting hormone, recently identified as strigolactone. Here, we report the cloning of SlCCD7 from tomato. As in other species, SlCCD7 encodes an enzyme capable of cleaving cyclic and acyclic carotenoids. However, the SlCCD7 protein has 30 additional amino acids of unknown function at its C terminus. Tomato plants expressing a SlCCD7 antisense construct display greatly increased branching. To reveal the underlying changes of this strong physiological phenotype, a metabolomic screen was conducted. With the exception of a reduction of stem amino acid content in the transgenic lines, no major changes were observed. In contrast, targeted analysis of the same plants revealed significantly decreased levels of strigolactone. There were no significant changes in root carotenoids, indicating that relatively little substrate is required to produce the bioactive strigolactones. The germination rate of Orobanche ramosa seeds was reduced by up to 90% on application of extract from the SlCCD7 antisense lines, compared with the wild type. Additionally, upon mycorrhizal colonization, C(13) cyclohexenone and C(14) mycorradicin apocarotenoid levels were greatly reduced in the roots of the antisense lines, implicating SlCCD7 in their biosynthesis. This work demonstrates the diverse roles of MAX3/CCD7 in strigolactone production, shoot branching, source-sink interactions and production of arbuscular mycorrhiza-induced apocarotenoids.

Ascertaining clonal fidelity of tissue culture raised plants of Bambusa balcooa Roxb. using inter simple sequence repeat markers

One of the major applications of plant tissue culture technology is that the regenerants are genetically identical to the mother plant. Although, of the various methods of in vitro propagation, axillary branching method is regarded as the safest, the possibility of genetic variation (somaclonal variation) cannot be ruled out even in this method. In the present study, inter simple sequence repeat (ISSR) marker assay was employed to validate the clonal fidelity of in vitro raised Bambusa balcooa plantlets multiplied by enhanced axillary proliferation up to 33 passages. Fifteen ISSR primers generated a total of 99 amplicons among the tissue-cultured progenies. Analysis of ISSR patterns revealed that the bands were shared by both the parent clump and the in vitro raised plants confirming the genetic stability in the latter.

Suppression of sorghum axillary bud outgrowth by shade, phyB and defoliation signalling pathways

In recent years, several genetic components of vegetative axillary bud development have been defined in both monocots and eudicots, but our understanding of environmental inputs on branching remains limited. Recent work in sorghum (Sorghum bicolor) has revealed a role for phytochrome B (phyB) in the control of axillary bud outgrowth through the regulation of Teosinte Branched1 (TB1) gene. In maize (Zea mays), TB1 is a dosage-dependent inhibitor of axillary meristem progression, and the expression level of TB1 is a sensitive measure of axillary branch development. To further explore the mechanistic basis of branching, the expression of branching and cell cycle-related genes were examined in phyB-1 and wild-type sorghum axillary buds following treatment with low-red : far-red light and defoliation. Although defoliation inhibited bud outgrowth, it did not influence the expression of sorghum TB1 (SbTB1), whereas changes in SbMAX2 expression, a homolog of the Arabidopsis (Arabidopsis thaliana) branching inhibitor MAX2, were associated with the regulation of bud outgrowth by both light and defoliation. The expression of several cell cycle-related genes was also decreased dramatically in buds repressed by defoliation, but not by phyB deficiency. The data suggest that there are at least two distinct molecular pathways that respond to light and endogenous signals to regulate axillary bud outgrowth.

A rapid in vitro multiplication system for commercial propagation of pharmaceutically important Cyclea peltata (Lam) Hook & Thoms. based on enhanced axillary branching

An efficient and quick in vitro propagation of Cyclea peltata by repeated subculture of nodal cuttings has been standardized. The nodal cuttings were cultured on Murashige and Skoog (MS) medium supplemented with various concentrations (0.5–7 mg/l) of 6-benzylaminopurine (BA) or kinetin (Kn) alone or in combination with indole-3-acetic acid (IAA; 0.5 mg/l) for culture initiation. Optimum response in terms of percent cultures responding, days to bud break and average shoot length was observed on MS medium fortified with 3 mg/l BA and 0.5 mg/l IAA. On this medium 90% cultures responded with 6.1 cm long unbranched vigorous solitary shoots in 45 days. The proliferated primary shoot emerged from the culture initiation step was incised into small nodal explants measuring a size of about 1.5 cm in length containing a single node and subcultured on multiplication medium supplemented with BA (0.5–7.0 mg/l) and IAA (0.5 mg/l). This process was repeated for another three subcultures (each of 45 days) to study the effect of subculturing on shoot multiplication. At the end of third passage 100% of nodal explants produced an average number of 14.2 healthy green shoots on MS medium supplemented with 3 mg/l BA and 0.5 mg/l IAA. The multiplied shoots were harvested and used for rooting on half-strength MS medium containing indole-3-butyric acid (IBA; 1–7 mg/l) or naphthalene acetic acid (NAA; 1–7 mg/l) for 45 days. The best rooting response was achieved on half-strength MS medium supplemented with 5 mg/l IBA. Here 96% cultures responded with an average number of 4.1 roots per shoot. Of the 40 plants transplanted to soil 28 survived (70%). This cost effective protocol will help the mass multiplication of C. peltata for commercial propagation.