Citrus Jambhiri Lush Research Articles

Agribusiness Prospects of Kachai Lemon Produced in Ukhrul District of Manipur, India: Status, Issues and Way Forward

Kachai lemon (Citrus jambhiri Lush.) is a variety of citrus, known for its rich nutritional and medicinal benefits due to its high flavonoid content and ascorbic acid. It has played a significant role in enhancing the socio-economic development of the tribal communities by generating income and employment opportunities. The Geographical Indication (GI) tag awarded to Kachai lemon has increased its commercial cultivation and market value. This recognition and growing attention have led to the emergence of agripreneurs and farmer producer organizations (FPOs) for production, value addition and marketing of Kachai lemon in an organized manner. The present study is aimed to compile the success stories, challenges of farmers and explore various Kachai lemon brands available on online platforms. A SWOT (Strength, Weakness, Opportunities and Threats) analysis on production and marketing of kachai lemon was performed. For the study, descriptive analysis was carried out to assess its potential, current status and challenges. The study suggests that in order to compete globally, it is crucial to develop strong post-harvest processing and marketing systems with support from agencies and institutions to provide valuable resources and market linkages to enhance efficiency and gain economic benefits.

Effect of Different Growing Media and Polybag Sizes on Seeds Germination & Root System of Rough Lemon (Citrus jambhiri Lush.) in Nursery Condition

Aims: The study aimed to evaluate the different growing media & polybag size for seedling growth of rough lemon rootstock. Study Design: The experiment used a Complete Randomized Design (CRD) with three replications. Place and Duration of Study: The study "Effect of Growing Media and Polybag Size on Rough Lemon Seedling Growth" took place in an insect-proof net house at CCS Haryana Agricultural University during the 2021-2022 academic year. Methodology: It comprising of 30 treatment combination i.e., ten different growing media: T1: garden soil; T2: soil: sand (1:1); T3: soil: sand: leafmold (1:1:1); T4: soil: cocopeat (1:1); T5: sand: cocopeat (1:1); T6: soil: sand: cocopeat (1:1:1); T7: soil: FYM (1:1); T8: sand: FYM (1:1); T9: soil: sand: FYM (1:1:1); T10: soil: sand: FYM: cocopeat (1:1:1:1), and the polybags S1: large (30 x 18 cm), S2: medium (21 x 18 cm) & S3: small size (17 x 16 cm) bags. Results: The study found that adding cocopeat to growing media (T4, T5, T6) resulted in earlier germination and higher rates (87-89%) Whereas, T4 to T10 media consists of either FYM, cocopeat for both resulted into more germination 87-89% as compared to controls (T1, T2, T3). The most of the root growth parameters i.e., diameter of tap root(2.45mm), fresh root weight (4.63g) & dry root weight (1.21g) & root: shoot ratio (0.61) improved with the media sand: FYM, soil: FYM, sand: soil: FYM, sand: soil: FYM: cocopeat in equal proportion. However, when media supplemented with cocopeat (T4, T5 & T6 ) also showed appreciable results as compared to soil, sand: soil & soil: sand: leafmold. However, sand: FYM, sand: cocopeat treatments were best in respect of number of primary roots (61.22), root volume (6.44ml). Whereas, sand: soil showed very poor results in all growth aspects. Among polybags sizes, large size (30 x 18cm) bags performed best in most of the germination & root growth parameters. Conclusion: Media enriched with organic matter outperformed sole media across all parameters, while larger polybags produced significantly better results than medium and small sizes.

Chemical profiling of citrus fruit-peel and stem extracts by modified UHPLC-TOF-MS and their antiplasmodial, antioxidant and antimicrobial activities

Citrus jambhiri (Lush) and paradisi (Macf.) have wide usage as food spices. Therefore, the presence of profiled phytochemicals, the antioxidant, antimicrobial and anti-plasmodial activities of the stem and fruit-peel extracts were studied. The citrus stem and fruit-peel extracts were analyzed using ultra-high-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UPLC-qTOF/MS) and anti-plasmodial activities against chloroquine-sensitive (CQS) strain of Plasmodium falciparum, antimicrobial against Gram-positive and Gram-negative bacterial strains as well as antioxidant potentials were determined in triplicate. Metabolite fingerprinting of C. jambhiri stem and C. paradisi fruit-peel extracts revealed rutamarin and spicatin respectively. The stem and fruit-peel of C. jambhiri showed moderate antimalarial activity. C. paradisi stem revealed a weak activity; however, C. paradisi fruit-peel showed no antimalarial activity (IC50 >100 μg/mL). C. paradisi and C. jambhiri extracts exhibited lower inhibition concentration (IC50: 3.0–7.0 µgmL−1), than ascorbic acid (IC50: 9.0 µgmL−1). C. paradisi stem DCM/methanol extract showed a reducing value of EC50: 8.00 µgmL−1 whereas reference compound used, (ascorbic acid) had EC50: 11.00 µgmL−1. Furthermore, C. jambhiri stem extract showed the best antimicrobial activity against the clinical isolates with high inhibition zones ≥ 15 mm. There is a paucity of information on the identification and importance of rutamarin and spicatin from Citrus extracts as a functional food ingredient with a promising compound implicated in the development of antimalarial drugs. Overall result however showed that the two citrus plants contain antioxidant constituents whereas C. jambhiri could be important source of antimicrobial agents. The study profiled two new compounds from the citrus extract and showed the potentials of the plant as source of antimalaria drug.

Monitoring dissociation of chimerism through real-time PCR and scanning electron microscopy following in planta transformation of rough lemon (Citrus jambhiri Lush.)

Monitoring dissociation of chimerism through real-time PCR and scanning electron microscopy following in planta transformation of rough lemon (Citrus jambhiri Lush.)

Optimization of explants, media, plant growth regulators and carbohydrates on callus induction and plant regeneration in Citrus jambhiri Lush.

Callus induction was attempted from the four explants viz. root, cotyledon, epicotyl, and leaf segments excised from in vitro raised seedlings of C. jambhiri. Among various MS media supplementations with growth regulators and carbohydrates, the maximum (95.50%) and the earliest (8.30 days) callogenesis was obtained in epicotyl segments, when cultured on MS medium supplemented with NAA (10.0 mgl-1) + BAP (1.0 mgl-1) + sucrose (8%). The modified MS (macro and micro-nutrients reduced to half) fortified with BAP (5.0 mgl-1) + GA3 (3.0 mgl-1) recorded maximum shoot regeneration (43.10%) from callus, with an average of 5.30 shoots per callus after 35.50 days of culturing. However, prolonged exposure to GA3 resulted in thin elongated shoots and leaves. The age of the callus substantially influenced the plant regeneration frequency. The potency of the callus to regenerate decreased significantly with an increase in the age of the callus. Shoot regeneration was recorded maximum (43.43%) in 60 days old calli, followed by 90 days old (30.48%) calli, whereas it was minimum (10.46%) in 150 days old calli. The maximum (79.50%) shoot proliferation was recorded in MS medium supplemented with BAP (1.0 mgl-1) + Kin (0.5 mgl-1) with an average of 5.06 shoots per culture. The MS medium fortified with NAA (1.0 mgl-1) + IBA (1.0 mgl-1) induced maximum (77.33%) rooting, with an average of 3.19 roots per shoot after 13.4 days of culturing. Rooted plants were hardened and survived the best (83.6%) on the potting mixture consisting of cocopeat + vermiculite + perlite (2:1:1).

Effects of salicylic acid application on germination, growth and development of rough lemon (Citrus jambhiri Lush.) under salt stress

Rough lemon (Citrus jambhiri Lush.) – RL, is used as a rootstock for citrus plants in saline conditions. NaCl causes an osmotic stress on plants mainly preventing the water uptake by the roots and thus reducing the plant growth. The objective of this study was to determine the effects of salicylic acid (SA) on germination of seeds and the growth and development of seedlings of RL rootstock under salt stress. For seed germination, a study was conducted in a completely randomized design in a 4 × 4 factorial scheme (SA at 0.00, 0.25, 0.50, or 1.00 mM and NaCl at 0, 50, 100, or 200 mM) with 4 repetitions, totaling 64 plots, of 25 seeds per plot. RL seeds were incubated in SA solutions for 24 h. Then, they were treated with NaCl-containing water in Petri dishes and incubated in the growth chamber at 25°C. For greenhouse experiment, a study was conducted in a randomized complete block in a 4 × 4 factorial scheme (SA at 0.0, 0.5, 1.0, or 2.0 mM and NaCl at 0, 50, 100, or 150 mM) with 3 repetitions, totaling 48 plots, of 2 plants per plot. Some morphological and physiological characteristics were determined. While germination time was extended, germination ratio and radicle extension were decreased in seeds under salt stress compared to control. Moreover, in these conditions the leaf membrane permeability and leaf falling were increased. In turn, plant height, diameter, root and shoot dry weight, leaf relative water content, and leaf chlorophyll were decreased in seedlings. Salt stress had negatively affected seed germination from 97.5% in control to 23.5% in 200 mM NaCl. However, SA treatments significantly decreased plant height to 67.8 cm in 2.0 mM compared to 80.1 cm in control in RL rootstock.

Impact of Different Potting Media on Growth of Rough Lemon (Citrus Jambhiri Lush)

Impact of Different Potting Media on Growth of Rough Lemon (Citrus Jambhiri Lush)

Zinc fertigation studies in Nagpur mandarin (Citrus reticulata) grown on black clay soil

A field experiment was conducted for four seasons (2013–17) on 12-year-old bearing Nagpur mandarin (Citrus reticulata Blanco) plants budded on rough lemon rootstock (Citrus jambhiri Lush) at ICAR-Central Citrus Research Institute, Nagpur, Maharashtra. Six zinc-based treatments, viz. S100 as soil application of ZnSO4 (100 g/plant), S200 as soil application of ZnSO4 (200 g/plant), S300 as soil application of ZnSO4 (300 g/plant, F100 as fertigation of ZnSO4 (100 g/plant), F200 as fertigation of ZnSO4 (200 g/plant) and F300 as fertigation of ZnSO4 (300 g/plant) with uniform doses of macronutrients (450 g N + 150 g P2O5 + 150 K2O/tree/year) were evaluated in a randomized block design (RBD) with four replications on a smectite rich alkaline black clay soil. Significantly higher concentration of Zn in index leaves was observed with fertigation treatments than soil application treatments with regard to fruit yield and quality. The fruit yield was observed significantly lower with soil application of ZnSO4 (11.5–13.95 tonnes/ha with S100-S300) than fertigation treatments (15.6–16.4 tonnes/ha with F100-F300). The fruit quality parameters were observed much higher with F200 followed by F300. On the other hand, the lowest acidity (0.82%) and higher TSS to acidity ratio (11.52) were observed with F200. Hence, the treatment F200, split through 4 pulses at 40 days interval from anthesis to the fruit development stage proved highly efficient for quality production of Nagpur mandarin in central India.

Assessing genetic diversity in rough lemon (Citrus jambhiri Lush) using morphological characters

Aim: To assess genetic diversity and relationship among important horticultural traits in rough lemon (Citrus jambhiri Lush). Methodology: The study was conducted at Punjab Agricultural University, Ludhiana on seventeen strains of rough lemon. Data was recorded for vegetative, fruiting and productivity parameters. Genetic variability and relationship among important horticultural traits were assessed by analyzing data with Microsoft Excel and Minitab 18 software. Results: There was wide variability among seventeen genotypes of rough lemon. Little difference between genotypic coefficient of variation (GCV) and phenotypic coefficient of variation (PCV) for majority of traits confirmed low environmental influence on the characters under study. High heritability with high to moderate genetic gain for fruit weight (69.49% and 36.50%), fruit length (95.24% and 40.62%), fruit diameter (93% and 32.23%), spine length (98.30% and 70.56%), acidity (95.07% and 40.18%) and fruit axis diameter (94.71% and 64.14%) suggested additive gene action and ability of improvement through selection. Strong character association of fruit number/tree and fruit axis diameter with yield revealed that these are the most important characteristics to be considered for effective selection in favour of high yield. Similarly, Principal Component Analysis showed high association between leaf lamina length and fruit size characters. Interpretation: This research suggests a high genetic diversity among the genotypes of rough lemon and valuable correlations were observed between different traits of economic significance. The findings of this study can be used in future crop improvement programmes for developing desirable varieties. Key words: Citrus jambhiri, Cluster analysis, Genetic diversity

Influence of basal vitamins, growth regulators, and explants on in vitro organogenesis from synthetic seeds of Citrus jambhiri Lush.

An efficient shoot and root organogenesis protocol has been developed from synseeds derived from various explants of Citrus jambhiri Lush. Optimum synseeds were developed using sodium alginate (0.5-0.75%) in 1.0% CaCl2 solution. Shoot organogenesis was examined under various basal vitamin mediums (MS Nitsch and MSB5) supplemented with various concentrations of adenine sulfate (ADS) and 6-benzyl amino purine (BAP) from different explants such as cotyledonary junction, shoot tip, and nodal explants. The synseed regeneration response ranged between 60-100% among the vitamins, cytokinins and explants used. The number of shoots per synseeds was higher (13.4) in MSB5-BAP (1.5 mg L-1), followed by 12.8 in MSN-ADS (2.0 mg L-1), 11.2 in MSN-ADS (1.0 mg L-1), and 10.8 in MSB5-ADS (1.0 mg L-1) from the synseeds developed using 0.75% sodium alginate. The mean number of roots per explant was higher (4.2) in ½MSN+IAA (1.0 mg L-1). Similarly, the mean root length was higher (5.2 cm) in ½MSN+IAA (0.5 mg L-1) followed by 4.2 cm in ½MSN+IAA (1.0 mg L-1). Regenerants derived from synseeds have shown no somaclonal variations, confirming that the plantlets are true-to-type to their parental progenies. The encapsulated plantlets showed >90% survivability while transferred at Kachai village, Manipur, India. The results of the present study encourage the use of various vitamin mediums and explants for large-scale propagation of C. jambhiri through synseeds.

Effect of chemical mutagens and Phytophthora culture filtrate on survival and plant regeneration from callus of Citrus jambhiri Lush.

Effect of chemical mutagens and Phytophthora culture filtrate on survival and plant regeneration from callus of Citrus jambhiri Lush.

​​Effect of Supplementation of Rough Lemon Juice, Amla Juice and Aloe Vera Gel on Performance of Broiler Chicken under Summer Season

Background: Herbal additives are rich in vitamin C, A and other biological active compounds and may act as a good alternative source to overcome heat stress in broiler chicken. Among herbal additives, rough lemon, amla and aloe vera were easily available during summer and also have an additional advantage of throw away price in comparison to synthetic feed additives available in market. Methods: In order to investigate the effect of supplementation of rough lemon (Citrus jambhiri Lush), amla (Emblica officinalis) and aloe vera (Aloe barbadensis Miller) on the performance of broiler chicken (Cobb strain) under summer season, 240-day old chicks were randomly distributed to four treatment groups (n=60). Basal diet (maize-soya based; T0) was same for all the groups except the supplementation of rough lemon juice, amla juice and aloe vera gel @ 2% in drinking water in T1, T2 and T3, respectively. Body weight gain, feed Intake, FCR were calculated during the experiment trial lasted for 42 days during June-July at F.V.Sc. and A.H., R.S. Pura, Jammu. The maximum and minimum temperature was recorded daily at 8:30 a.m. and 2:30 p.m. by using digital maximum minimum thermometer. Result: Body weight gain and FCR was found superior in T3 (P greater than 0.05) than T2 and T1 and was inferior in T0. Weekly water intake (ml/bird) was lower in T2 and T3. Benefit cost ratio was highest in T3 (1.313) followed by T2 (1.271), T1 (1.231) and least in T0 (1.071). It may be concluded that aloe vera gel supplementation resulted in better broiler performance followed by amla juice and rough lemon juice supplemented groups.

The Number of Bacteria Colonies in Carp Fish (Cyprinus carpio) After Administration of Lime (Citrus aurantifolia) and Orange Extract (Citrus jambhiri Lush.)

Fish is one of the most commonly consumed protein sources. Fish are at risk as a medium for the growth of microbial contaminants that can cause foodborne diseases. This disease can cause diarrhea and even death.Contaminant microbes can be found in foods consumed raw, such as Naniura, a typical Batak cuisine in the form of raw carp. Various natural spices such as lime and orange also have potential as antimicrobials. This study aimed to determine the antimicrobial potential of lime and orange juice in fresh carp samples. This study used extracts of lime and orange with concentrations of 6.25%, 12.5%, 25%, and 50%, respectively. The sample used is carp meat. The bacteria present in the sample were counted using the Total Plate Count (TPC) method before and after lime and lime extract administration. The results showed that before administering the two extracts, the total colonies in the sample could not be counted. The antimicrobial effect of orange juice extract began to be seen at a concentration of 12.5%, while in lime, it was only seen at a concentration of 50%. From the results of this study, it was concluded that orange juice extract had a better antimicrobial effect than lime.

Transcriptome Analysis of Plenodomus tracheiphilus Infecting Rough Lemon (Citrus jambhiri Lush.) Indicates a Multifaceted Strategy during Host Pathogenesis.

Simple SummaryThe cultivation of the lemon is strongly impacted by mal secco, a disease that causes huge losses in yield every year. In this work, we have identified, retrieved, and classified genes that may play a crucial role in the onset and progression of the disease. Understanding the function of these genes will increase knowledge of the processes involving the mode of action of necrotrophic fungi during pathogenesis. Our results may be relevant to help identify sustainable field treatments to cope with disease diffusion and to provide direction into possible biotechnological approaches to generate resistant lemon plants.The causal agent of mal secco disease is the fungus Plenodomus tracheiphilus, mainly affecting lemon tree survival in the Mediterranean area. Using a fully compatible host-pathogen interaction, the aim of our work was to retrieve the fungus transcriptome by an RNA seq approach during infection of rough lemon (Citrus jambhiri Lush.) to identify crucial transcripts for pathogenesis establishment and progression. A total of 2438 clusters belonging to P. tracheiphilus were retrieved and classified into the GO and KEGG categories. Transcripts were categorized mainly within the “membrane”, “catalytic activity”, and “primary metabolic process” GO terms. Moreover, most of the transcripts are included in the “ribosome”, “carbon metabolism”, and “oxidative phosphorylation” KEGG categories. By focusing our attention on transcripts with FPKM values higher than the median, we were able to identify four main transcript groups functioning in (a) fungus cell wall remodeling and protection, (b) destroying plant defensive secondary metabolites, (c) optimizing fungus development and pathogenesis, and (d) toxin biosynthesis, thus indicating that a multifaceted strategy to subdue the host was executed.

Evaluation of Integrated Management Approaches against Citrus Nematode (Tylenchulus semipenetrans) in Pakistan

Present study was conducted to explore the nematicidal potential of different synthetic chemicals, biopesticides and antagonists against citrus decline. Effect of eleven chemicals, seven biopesticides and two antagonists on juvenile mortality was evaluated in vitro. Four concentrations (2S, S, S/2 and S/4) of each chemical were prepared on the basis of recommended dose for each chemical. Juvenile mortality of citrus nematodes was calculated after 24, 48 and 72 hour of exposure to chemicals. Rugby expressed maximum percentage of juvenile mortality at 2S, S, S/2 and S/4 concentration. Mortality percentage observed by Rugby, Furadan, Match and Cartap after 72 h at 2S concentration was (100, 100, 95.44, 88.23%) whereas at S/4 concentration mortality percentage was (76.32, 81.18, 62.15, 55.28%) respectively. Among biopesticides maximum percentage mortality observed by Proclaim and Cure after 72 hours at 2S concentration was (83.87, 80.44%) while at S/4 concentration it was (59.87, 57.38%) respectively. Cultural filtrates of two antagonist Trichoderma harzianum and Trichoderma viridi were evaluated at S, S/2 and S/4 concentration under lab conditions. Maximum mortality (88.42%) was observed when both antagonists were applied in combined treatment at S concentration after 72 hours of exposure. Two best performing chemicals, one biopesticide and one antagonist were evaluated under greenhouse against T. semipenetrans on Citrus jambhiri Lush (rough lemon) and their effect on plant growth and nutrient uptake was measured. Nitrogen uptake was measured by following micro Kjeldahl method. For phosphorus and potassium uptake absorbance of samples at 720 nm was measured with the help of spectrophotometer. Phosphorus was calculated by comparing standard curve already prepared while potassium uptake was assessed by flame photometer method after digestion. All the treatments were found significantly effective against citrus nematode but maximum plant height, stem diameter, number of leaves, root length, root weight, shoot weight, and number of feeder roots (90 cm, 2.4 cm, 102, 48.3 g, 12.1 g, 48.8 g and 70) as compared to control when Rugby, Furadan, Proclaim and T. harzianum were applied in combination. Similarly maximum uptake of Nitrogen (3.14) Phosphorous (2.44%) and Potassium (1.95%) was observed when all the treatments were applied in combination. The results of present study will be helpful in selecting the suitable chemicals for growers having problems of citrus nematodes in orchards.

Quantification of phenolic constituents and bioactive properties of callus and leaf tissue of Citrus jambhiri lush

The chemical constituents of callus and leaf tissues of Citrus jambhiri Lush were extracted in methanol, ethanol and chloroform and the extracts were estimated for their total phenolic and flavonoid contents (TPC and TFC).Evaluation of antioxidant and antibacterial potential of callus and leaf extracts revealed that methanol extracts exhibited higher DPPH radical scavenging activity (radical scavenging index 4.55 of leaf extract and 9.09 of standard BHA).Further, these extracts were most effective against pathogenic bacteria (with inhibition zone diameters 8.0 and 8.9 mm against Escherichia coli and Salmonella typhi, respectively). Leaf extracts were found to contain slightly higher amounts of TPC and TFC and hence, higher antioxidant and antibacterial activities as compared to callus extracts.

Scheduling of nutrient doses for rough lemon (Citrus jambhiri) rootstock under containerized primary and secondary nursery

The study was carried out to investigate and standardize the nutritional requirement of commonly used roughlemon (Citrus jambhiri Lush) citrus rootstock for raising commercial primary and secondary containerized nursery.The aim of the study was also to reduce fairly high (2 years) time required traditionally for seedling production incitrus nurseries and to provide healthy seedlings. Two separate experiments, comprising different nutrient doseswere conducted under primary and secondary containerized nursery. The results revealed that dry weight of all theplant parts and number of leaves was maximum with the application of 40+25+25 g NPK. Uptake of all the macronutrients,viz. N (31.2 mg), P (1.33 mg) and K (27.7 mg) by the plants was also highest under this treatment. Whileplant height and leaf chlorophyll contents were better in 20+10+10g NPK as compared to control. In secondarynursery, application of 4+2+2 g NPK recorded maximum plant height, stem girth, budding success rate (95.8%) andsignificantly reduced the time required for budding. Although availability of major nutrients was significantly highestin 60+25+25 g NPK treatment, it could not encourage the growth of the plants and induced plant mortality. In primarynursery, application of 40+25+25 g NPK and in secondary nursery, application 4+2+2 g NPK potting mixture wereproved to be the superior nutrient dosage for rough lemon rootstock.

Morpho-genetic diversity in zygotic populations derived from Rough lemon (Citrus jambhiri Lush.) and their tolerance against Phytophthora

Morpho-genetic diversity in zygotic populations derived from Rough lemon (Citrus jambhiri Lush.) and their tolerance against Phytophthora

High efficient de novo root-to-shoot organogenesis in Citrus jambhiri Lush.: Gene expression, genetic stability and virus indexing.

A protocol for high-frequency direct organogenesis from root explants of Kachai lemon (Citrus jambhiri Lush.) was developed. Full-length roots (~3 cm) were isolated from the in vitro grown seedlings and cultured on Murashige and Skoog basal medium supplemented with Nitsch vitamin (MSN) with different concentrations of cytokinin [6-benzylaminopurine, (BAP)] and gibberellic acid (GA3). The frequency of multiple shoot proliferation was very high, with an average of 34.3 shoots per root explant when inoculated on the MSN medium supplemented with BAP (1.0 mg L-1) and GA3 (1.0 mg L-1). Optimal rooting was induced in the plantlets under half strength MSN medium supplemented with indole-3-acetic acid (IAA, 0.5-1.0 mg L-1). IAA induced better root structure than 1-naphthaleneacetic acid (NAA), which was evident from the scanning electron microscopy (SEM). The expressions of growth regulating factor genes (GRF1 and GRF5) and GA3 signaling genes (GA2OX1 and KO1) were elevated in the regenerants obtained from MSN+BAP (1.0 mg L-1)+GA3 (1.0 mg L-1). The expressions of auxin regulating genes were high in roots obtained in ½ MSN+IAA 1.0 mg L-1. Furthermore, indexing of the regenerants confirmed that there was no amplicons detected for Huanglongbing bacterium and Citrus tristeza virus. Random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) markers detected no polymorphic bands amongst the regenerated plants. This is the first report that describes direct organogenesis from the root explant of Citrus jambhiri Lush. The high-frequency direct regeneration protocol in the present study provides an enormous significance in Citrus organogenesis, its commercial cultivation and genetic conservation.

Effect of different growth regulators on in vitro regeneration of cotyledon explants of rough lemon (Citrus jambhiri Lush.) via callus induction

In order to establish in vitro protocols for raising aseptic cultures of cotyledon segments excised from five-week-old in vitro raised seedlings were used as explant material. Among surface sterilants, mercuric chloride 0.1% for three minutes provided least rate of contamination (24%) and the maximum explant survival (76%). The highest per cent culture establishment of 97.79% was obtained in cotyledon explants on Murashige and Skoog (MS) containing 2,4-D (3.0 mg/l) and maximum callus induction (95.41%) was observed on MS medium supplemented with 2,4-D (2.0 mg/l). Among different plant growth regulator treatments, maximum callus regeneration of 82.19% was observed on MS medium supplemented with BAP (3.0 mg/l) whereas, minimum callus regeneration (4.33%) was found in MS medium supplemented with BAP (3.0 mg/l) in combination with IAA (0.75 mg/l) and malt extract (500 mg/l). Maximum rooting response (96.00%) was recorded on MS (half strength) medium with NAA (2.0 mg/l) and was at par with rooting percentage of 92.00% on MS (half strength) medium supplemented with NAA (1.5 mg/l). The plantlets were successfully acclimatized in different potting mixtures and highest survival rate (91.35%) was achieved in potting mixture containing garden soil with sand and farmyard manure (1:1:1).