Physiological and transcriptome analysis of changes in endogenous hormone and sugar content during the formation of tender asparagus stems.

Asparagus is a nutritionally dense stem vegetable whose growth and development are correlated with its quality and yield. To investigate the dynamic changes and underlying mechanisms during the elongation and growth process of asparagus stems, we documented the growth pattern of asparagus and selected stem segments from four consecutive elongation stages using physiological and transcriptome analyses. Notably, the growth rate of asparagus accelerated at a length of 25 cm. A significant decrease in the concentration of sucrose, fructose, glucose, and additional sugars was observed in the elongation region of tender stems. Conversely, the levels of auxin and gibberellins(GAs) were elevated along with increased activity of enzymes involved in sucrose degradation. A significant positive correlation existed between auxin, GAs, and enzymes involved in sucrose degradation. The ABA content gradually increased with stem elongation. The tissue section showed that cell elongation is an inherent manifestation of stem elongation. The differential genes screened by transcriptome analysis were enriched in pathways such as starch and sucrose metabolism, phytohormone synthesis metabolism, and signal transduction. The expression levels of genes such as ARF, GA20ox, NCED, PIF4, and otherswere upregulated during stem elongation, while DAO, GA2ox, and other genes were downregulated. The gene expression level was consistent with changes in hormone content and influenced the cell length elongation. Additionally, the expression results of RT-qPCR were consistent with RNA-seq. The observed variations in gene expression levels, endogenous hormones and sugar changes during the elongation and growth of asparagus tender stems offer valuable insights for future investigations into the molecular mechanisms of asparagus stem growth and development and provide a theoretical foundation for cultivation and production practices.

Environmental footprint of critical agro-export products in the Peruvian hyper-arid coast: A case study for green asparagus and avocado.

Peru has become one of the world's main agricultural hubs for a wide range of fruits and vegetables. Two of these products, avocado and green asparagus, have raised attention in recent years in the international scene from an environmental perspective due to the high amounts of water they require, as well as the long air and marine freighting distances to export these products to Europe, Asia or the US. Consequently, the aim of the current study was to perform an environmental assessment of these two products using two life-cycle methods: carbon and water footprint. For the latter, water scarcity, acidification, eco-toxicity and eutrophication impact categories have been selected for assessment. Inventory data were gathered from six different companies located in different regions of the hyper-arid Peruvian coast. The results report that the products are not carbon intensive and are in line with other similar plant-based products. Conversely, the hyper-arid conditions of the cultivation sites require a large volume of groundwater to fulfill the needs of the crops. Interestingly, even though this may lead to overexploitation of groundwater resources in the absence of appropriate management policies, the low mobility of pollutants, namely pesticides, constitutes a natural barrier to protect the degradation of natural water bodies. Similarly, highly technified irrigation systems have allowed minimizing the amounts of water used per hectare. In conclusion, results from this study may be useful in more concise environmental assessment studies on food products and diets, considering the consumption of these Peruvian products in many countries in the world. Furthermore, results are also important at regional level since they depict the carbon and water performance of these products and can also be accompanied by cross-cutting certification schemes, including Product Environmental Footprint Category Rules Guidance.

Changes in the primary and secondary metabolome of male green asparagus (Asparagus officinalis L.) as modulated by sequential harvesting.

Rising temperature and solar radiation drive the mobilization and depletion of crown-deposited metabolites harnessed for asparagus spear regeneration during the spring harvest season. We presently examined how successive same-season harvests impact the sensory, nutritive and bioactive composition of select green asparagus genotypes. Soluble carbohydrates were analyzed by HPLC-RI, organic acids and polyphenols by HPLC-DAD and metals by ion chromatography. Higher sugars and lower acids accentuated sweetness and lower polyphenols contributed to reduced astringency at the beginning of the harvest season. This trend was reversed as the season advanced and spear sensory quality was compromised by declining sugars and rising acids; however, functional quality improved as antioxidant capacity increased along with the concentrations of quercetin-3-O-rutinoside (rutin) and ascorbic acid. The compositional changes presently described were uniform across all genotypes examined and thus contribute toward our understanding of seasonal variation in the sensory and functional quality of this acclaimed health-promoting product.

A cytogenetic analysis of male meiosis in Asparagus officinalis.

Asparagus (Asparagus officinalis) has several traits that make it a useful model for cytogenetic studies, however, few studies of the meiosis process have been made in asparagus. Here, we present in detail an atlas of male meiosis in asparagus, from preleptotene to telophase II. The meiosis process in asparagus is largely similar to those of the well-characterized model plants Arabidopsis thaliana, Zea mays, and Oryza sativa. However, most asparagus prophase I meiotic chromosomes show a strongly aggregated morphology, and this phenotype persists through the pachytene stage, highlighting a property in the control of chromosome migration and distribution in asparagus. Further, we observed no obvious banding of autofluorescent dots between divided nuclei of asparagus meiocytes, as one would expect in Arabidopsis. This description of wild-type asparagus meiosis will serve as a reference for the analyses of meiotic mutants, as well as for comparative studies among difference species. Abbreviations: DAPI: 4',6-diamidino-2-phenylindole; FISH: fluorescence in situ hybridization; PBS: phosphate-buffered saline; PMC: pollen mother cell; SEM: Scanning Electron Microscope.

Involvement of an autotoxic compound in asparagus decline.

Asparagus (Asparagus officinalis L.) is a widely cultivated perennial veritable and can be harvested more than ten years. However, the crop quality and yield decline after a few year's cultivation, which is called "asparagus decline". Even though those asparagus plants were replaced with new young asparagus plants, the productivity and quality of the crop remain relatively low, which is known as a "asparagus replant problem". One of the possible reasons for "asparagus decline" and "asparagus replant problem" is thought to be autotoxicity of asparagus. However, the compounds involved in the autotoxicity is not clear. The objective of this study was therefore to determine the potential role of autotoxicity in the "asparagus decline" and "asparagus replant problem". An aqueous methanol extract of 10-year-asparagus-cultivated soils inhibited the growth of asparagus seedlings and other two test plants with concentration dependent manner. The result confirmed that the asparagus soils have autotoxic activity. The extract was then purified by several chromatographies with monitoring the inhibitory activity and a potent growth inhibitory substance causing the autotoxic effect was isolated. The chemical structures of the compound was determined by spectral data to be trans-cinnamic acid. trans-Cinnamic acid inhibited the growth of asparagus seedlings at concentrations greater than 10 µM. The concentrations required for 50% growth inhibition of asparagus (IC50) were 24.1-41.6 µM. trans-Cinnamic acid accumulated 174 µM in the 10-year-asparagus-cultivated soils, which may be enough levels to cause the growth inhibition on asparagus considering its IC50 value. Therefore, trans-cinnamic acid may contribute to the autotoxic effect of asparagus soils, and may be in part responsible for "asparagus decline" and "asparagus replant problem".

Influence of different cultivation systems on bioactivity of asparagus.

Evaluation of functional and bioactive compounds of Asparagus officinalis L. growing in conventional and organic greenhouse and conventional open - field farming was the aim of this research. Polyphenols in cladodes grown conventionally were higher than organic. Flavonoids and carotenoids were the highest in cladodes in open field. Organic spears were richer in total phenolics (+6.9% and +19.1%) and flavonoids (+4.7% and 16.8%) and showed higher in vitro averages of antioxidant activities by three radical scavenging assays (+12.5% and+ 22.2%) than conventional. Partial differentiation of organically and conventionally grown samples was proved by multivariate statistics. The binding properties of polyphenols to HSA were relatively high in comparison with other plants. A strong positive correlation of binding properties and bioactivity of asparagus was estimated. All new found aspects for the first time lead to recommendation of inclusion of all investigated asparagus plants into the human diet in a wider scale.

Comparative transcriptome analysis reveals differentially expressed genes associated with sex expression in garden asparagus (Asparagus officinalis).

BACKGROUND: Garden asparagus (Asparagus officinalis) is a highly valuable vegetable crop of commercial and nutritional interest. It is also commonly used to investigate the mechanisms of sex determination and differentiation in plants. However, the sex expression mechanisms in asparagus remain poorly understood. RESULTS: De novo transcriptome sequencing via Illumina paired-end sequencing revealed more than 26 billion bases of high-quality sequence data from male and female asparagus flower buds. A total of 72,626 unigenes with an average length of 979 bp were assembled. In comparative transcriptome analysis, 4876 differentially expressed genes (DEGs) were identified in the possible sex-determining stage of female and male/supermale flower buds. Of these DEGs, 433, including 285 male/supermale-biased and 149 female-biased genes, were annotated as flower related. Of the male/supermale-biased flower-related genes, 102 were probably involved in anther development. In addition, 43 DEGs implicated in hormone response and biosynthesis putatively associated with sex expression and reproduction were discovered. Moreover, 128 transcription factor (TF)-related genes belonging to various families were found to be differentially expressed, and this finding implied the essential roles of TF in sex determination or differentiation in asparagus. Correlation analysis indicated that miRNA-DEG pairs were also implicated in asparagus sexual development. CONCLUSIONS: Our study identified a large number of DEGs involved in the sex expression and reproduction of asparagus, including known genes participating in plant reproduction, plant hormone signaling, TF encoding, and genes with unclear functions. We also found that miRNAs might be involved in the sex differentiation process. Our study could provide a valuable basis for further investigations on the regulatory networks of sex determination and differentiation in asparagus and facilitate further genetic and genomic studies on this dioecious species.

MYB transcription factor gene involved in sex determination in Asparagus officinalis.

Dioecy is a plant mating system in which individuals of a species are either male or female. Although many flowering plants evolved independently from hermaphroditism to dioecy, the molecular mechanism underlying this transition remains largely unknown. Sex determination in the dioecious plant Asparagus officinalis is controlled by X and Y chromosomes; the male and female karyotypes are XY and XX, respectively. Transcriptome analysis of A. officinalis buds showed that a MYB-like gene, Male Specific Expression 1 (MSE1), is specifically expressed in males. MSE1 exhibits tight linkage with the Y chromosome, specific expression in early anther development and loss of function on the X chromosome. Knockout of the MSE1 orthologue in Arabidopsis induces male sterility. Thus, MSE1 acts in sex determination in A. officinalis.

Antiradical capacity and polyphenol composition of asparagus spears varieties cultivated under different sunlight conditions.

BACKGROUND: Asparagus officinalis has a high nutritional value. Asparagus is rich in a number of bioactive compounds, mainly flavonoids (quercetin), glutathione, vitamin C, vitamin E, fructans (inulin and fructooligosaccharides) and phytosterols (b-sitosterol). These compounds may play an important role in human health. The purpose of this study was to examine the antioxidant potential and polyphenol composition of white, pale-colored and green asparagus spears of different cultivars. METHODS: Investigations were conducted on different asparagus spear extracts. The study included three colors of asparagus (white, pale-colored and green) from five different cultivars subjected to the ethanol extraction procedure. Total phenolic content was also determined by the Folin-Ciocalteu method. Polyphenol (phenolic acids and flavonols) composition was estimated using the HPLC method. The antioxidant properties of extracts were examined using DPPH, ABTS and metal ion chelating assays. RESULTS: The highest contents of phenolic and flavonoids were observed in green asparagus from Grolim and the lowest in pale-colored asparagus from Gyjmlin. It was found that both the color of asparagus and the cultivar had a significant effect on the composition of phenolic acid and flavonols. Radical scavenging activity toward DPPH• and ABTS was highest for green asparagus cv. Grolim and Eposs. The greatest number of Fe ions was chelated by samples of green asparagus cv. Grolim and Huchel's Alpha and pale-colored asparagus cv. Huchel's Alpha. CONCLUSIONS: It was shown that the antioxidant activity of asparagus spears measured by antiradical and chelating activity test depends on variety and color. The highest activity was found in green asparagus and the lowest was identified in white asparagus extracts. It has also been clarified that changes in flavonol and phenolic acid composition and increases in their diversity depends on growing with sunlight and variety. Asparagus can provide a valuable source of phenolic compounds in the human diet.

Isolation of differentially expressed sex genes in garden asparagus using suppression subtractive hybridization.

Garden asparagus (Asparagus officinalis L.) is a dioecious species whose male and female flowers are found in separate unisexual individuals. A region called the M-locus, located on a pair of homomorphic sex chromosomes, controls sexual dimorphism in asparagus. To date, no sex determining gene has been isolated from asparagus. To identify more genes involved in flower development in asparagus, subtractive hybridization library of male flowers in asparagus was constructed by suppression subtraction hybridization. A total of 107 expressed sequence tags (ESTs) were identified. BLASTX analysis showed that the library contained several genes that could be related to flower development. The expression patterns of seven selected genes believed to be involved in the development of asparagus male flower were further analyzed by semi-quantitative or real-time reverse-transcription polymerase chain reaction (RT-PCR). Results showed that AOEST4-5, AOEST12-40, and AOEST13-38 were strongly expressed in the male flower stage, whereas no transcript level of AOEST13-38 was detected in the female flower stage. The expression levels of AOEST13-87, AOEST13-92, AOEST13-40, and AOEST18-87 in the male flower stage were also higher than those in the female flower stage, although these transcripts were also expressed in other tissues. The identified genes can provide a strong starting point for further studies on the underlying molecular differences between the male and female flowers of asparagus.

Sex-biased gene expression in dioecious garden asparagus (Asparagus officinalis).

Sex chromosomes have evolved independently in phylogenetically diverse flowering plant lineages. The genes governing sex determination in dioecious species remain unknown, but theory predicts that the linkage of genes influencing male and female function will spur the origin and early evolution of sex chromosomes. For example, in an XY system, the origin of an active Y may be spurred by the linkage of female suppressing and male promoting genes. Garden asparagus (Asparagus officinalis) serves as a model for plant sex chromosome evolution, given that it has recently evolved an XX/XY sex chromosome system. In order to elucidate the molecular basis of gender differences and sex determination, we used RNA-sequencing (RNA-Seq) to identify differentially expressed genes between female (XX), male (XY) and supermale (YY) individuals. We identified 570 differentially expressed genes, and showed that significantly more genes exhibited male-biased than female-biased expression in garden asparagus. In the context of anther development, we identified genes involved in pollen microspore and tapetum development that were specifically expressed in males and supermales. Comparative analysis of genes in the Arabidopsis thaliana, Zea mays and Oryza sativa anther development pathways shows that anther sterility in females probably occurs through interruption of tapetum development before microspore meiosis.

Bioactive compounds, antioxidant and binding activities and spear yield of Asparagus officinalis L.

The aim of this investigation was to find a proper harvesting period and establishing fern number, which effects the spear yield, bioactive compounds and antioxidant activities of Asparagus officinalis L. Spears were harvested at 2, 4, and 6 weeks after sprouting. Control for comparison was used without harvest. Spears and total yield increased with prolonged spear harvest period. In harvest of 6 weeks long optimum spear yield was the highest and fern numbers were 5 ~ 8. Bioactive compounds (polyphenols, flavonoids, flavanols, tannins and ascorbic acid) and the levels of antioxidant activities by ferric-reducing/antioxidant power (FRAP) and cupric reducing antioxidant capacity (CUPRAC) assays in asparagus ethanol extracts significantly differed in the investigated samples and were the highest at 6 weeks harvest period (P < 0.05). The first and the second segments from the tip significantly increased with the increase of catalase (CAT). It was interesting to investigate in vitro how human serum albumin (HSA) interacts with polyphenols extracted from investigated vegetables. Therefore the functional properties of asparagus were studied by the interaction of polyphenol ethanol extracts with HSA, using 3D- FL. In conclusion, antioxidant status (bioactive compounds, binding and antioxidant activities) improved with the harvesting period and the first segment from spear tip. Appropriate harvesting is effective for higher asparagus yield and its bioactivity.

Effect of chitosan and its derivatives as antifungal and preservative agents on postharvest green asparagus.

The antifungal activity and effect of high-molecular weight chitosan (H-chitosan), low-molecular weight chitosan (L-chitosan) and carboxymethyl chitosan (C-chitosan) coatings on postharvest green asparagus were evaluated. L-chitosan and H-chitosan efficiently inhibited the radial growth of Fusarium concentricum separated from postharvest green asparagus at 4 mg/ml, which appeared to be more effective in inhibiting spore germination and germ tube elongation than that of C-chitosan. Notably, spore germination was totally inhibited by L-chitosan and H-chitosan at 0.05 mg/ml. Coated asparagus did not show any apparent sign of phytotoxicity and maintained good quality over 28 days of cold storage, according to the weight loss and general quality aspects. Present results inferred that chitosan could act as an attractive preservative agent for postharvest green asparagus owing to its antifungal activity and its ability to stimulate some defense responses during storage.

Characterization of Fusarium isolates from asparagus fields in southwestern Ontario and influence of soil organic amendments on Fusarium crown and root rot.

Fusarium crown and root rot (FCRR) of asparagus has a complex etiology with several soilborne Fusarium spp. as causal agents. Ninety-three Fusarium isolates, obtained from plant and soil samples collected from commercial asparagus fields in southwestern Ontario with a history of FCRR, were identified as Fusarium oxysporum (65.5%), F. proliferatum (18.3%), F. solani (6.4%), F. acuminatum (6.4%), and F. redolens (3.2%) based on morphological or cultural characteristics and polymerase chain reaction (PCR) analysis with species-specific primers. The intersimple-sequence repeat PCR analysis of the field isolates revealed considerable variability among the isolates belonging to different Fusarium spp. In the in vitro pathogenicity screening tests, 50% of the field isolates were pathogenic to asparagus, and 22% of the isolates caused the most severe symptoms on asparagus. The management of FCRR with soil organic amendments of pelleted poultry manure (PPM), olive residue compost, and fish emulsion was evaluated in a greenhouse using three asparagus cultivars of different susceptibility in soils infested with two of the pathogenic isolates (F. oxysporum Fo-1.5 and F. solani Fs-1.12). Lower FCRR symptom severity and higher plant weights were observed for most treatments on 'Jersey Giant' and 'Grande' but not on 'Mary Washington'. On all three cultivars, 1% PPM consistently reduced FCRR severity by 42 to 96% and increased plant weights by 77 to 152% compared with the Fusarium control treatment. Populations of Fusarium and total bacteria were enumerated after 1, 3, 7, and 14 days of soil amendment. In amended soils, the population of Fusarium spp. gradually decreased while the population of total culturable bacteria increased. These results indicate that soil organic amendments, especially PPM, can decrease disease severity and promote plant growth, possibly by decreasing pathogen population and enhancing bacterial activity in the soil.

The development of the asparagus miner (Ophiomyia simplex Loew; Diptera: Agromyzidae) in temperate zones: a degree-day model.

BACKGROUND: The asparagus miner is a putative vector of Fusarium spp., which have been implicated in globally declining asparagus production. Growers currently apply broad-spectrum insecticides for the asparagus miner, but lack management guidelines for adequately controlling the pest. Our aims were (1) to determine the lower developmental threshold of the asparagus miner, (2) develop and validate a degree-day model describing its phenology, and (3) create a developmental time budget for the asparagus miner to help guide growers' management decisions. RESULTS: We found that the lower developmental threshold for the asparagus miner was 12.1 °C, and that the phenology of the asparagus miner could be reliably predicted over the course of a two-year study. Predictions from the model match well with previously published information on the bionomics of the asparagus miner, but fit better for sampling data collected from the midwestern and eastern United States than for the United Kingdom. The life cycle of the asparagus miner likely requires between 1500 and 2000 degree-days to complete; the longest developmental time requirement was for the pupal stagen CONCLUSION: This study provides tools for the targeted management of the asparagus miner by offering a degree-day model that may be used to predict its life stages in the north-eastern United States.

DAMAGE RESEARCH WITH P. PENETRANS IN ASPARAGUS PLANTS.

During cultivation of asparagus plants growth can be inhibited and yield can be reduced by plant-parasitic nematodes. Plant raising companies assume that the root lesion nematode (Pratylenchus penetrans) can cause severe yield loss in asparagus plants. However quantitative information about yield reduction in relation to the degree of infestation of this nematode species in the field is lacking. Research was done in The Netherlands by Applied Plant Research (part of Wageningen University and Research Centre) to determine the maximum degree of yield loss of asparagus plants at high infestation levels of P. penetrans and to establish the height of the tolerance limit for this nematode species. Also was investigated whether a field application of a granular nematicide could prevent or reduce yield loss caused by P. penetrans. Research was done in the field at sandy soils at the PPO location near Vredepeel in The Netherlands over a period of two years. In the first year the most suitable field was selected and on this field different infestation levels of P. penetrans were created. In the second year asparagus was cultivated and plant yield (number and quality of deliverable plants and financial yield) was calculated. At high infestation levels of Pratylenchus penetrans maximum yield loss was 12% (which can mean a financial loss of 7.000 C per ha). Yield started to decrease at very low infestation levels of P. penetrans and no statistical reliable tolerance limit could be calculated. Field application of 40 kg per ha of Vydate 10 G just before sowing of asparagus, could almost completely prevent yield loss caused by P. penetrans. After harvest infestation levels of P. penetrans were much lower than could be expected if asparagus was a non-host for this nematode species. In this paper therefore it is suggested that asparagus plants are (actively) controlling P. penetrans.

Patterns of spatial and temporal distribution of the asparagus miner (Diptera: Agromyzidae): implications for management.

The asparagus miner is an obligatory feeder on asparagus and a putative vector for pathogenic fungi implicated in the early decline of asparagus fields. To date, the distribution of the asparagus miner over space and time is poorly understood. Our study evaluated the spatial and temporal pattern of adult asparagus miners in commercial asparagus fields in Michigan in 2011 and 2012. We sampled adults and damage weekly during the growing season using yellow sticky traps outside, at the edge, and inside commercial fields. Yellow sticky traps at each trapping location were placed at the canopy and ground level to determine vertical distribution of adults. During the first generation, adults were more evenly distributed throughout the field. In the second generation, adults were more commonly found on the edge of the field. Overall, there was a greater percent of mining damage near the edge of the field. Additionally, three times as many asparagus miners were found in the canopy compared with ground-level traps. There were 12 times as many asparagus miner adults on edges bordered by another asparagus field than on ones bordered by forest. Taken together, our results indicate that while asparagus miner management in the beginning of the growing season should focus on the entire field, in the latter half of the season, growers could save money and resources by targeting miner adults at the edges of fields. Finally, conserving the remaining naturally forested landscape and planting borders of trees may help ameliorate pest pressure in asparagus fields.

Comparative analysis of gene expression by microarray analysis of male and female flowers of Asparagus officinalis.

To identify rapidly a number of genes probably involved in sex determination and differentiation of the dioecious plant Asparagus officinalis, gene expression profiles in early flower development for male and female plants were investigated by microarray assay with 8,665 probes. In total, 638 male-biased and 543 female-biased genes were identified. These genes with biased-expression for male and female were involved in a variety of processes associated with molecular functions, cellular components, and biological processes, suggesting that a complex mechanism underlies the sex development of asparagus. Among the differentially expressed genes involved in the reproductive process, a number of genes associated with floral development were identified. Reverse transcription-PCR was performed for validation, and the results were largely consistent with those obtained by microarray analysis. The findings of this study might contribute to understanding of the molecular mechanisms of sex determination and differentiation in dioecious asparagus and provide a foundation for further studies of this plant.

Ecological investigation of three geophytes in the Deltaic Mediterranean coast of Egypt.

This study was conducted to investigate the ecological features of three geophytes namely Asparagus stipularis, Cyperus capitatus and Stipagrostis lanata which grow naturally in the Nile Delta coast of Egypt. C. capitatus and S. lanata are growing in non-saline sandy soils and can tolerate drought stress while, A. stipularis is growing in saline and non-saline sandy and calcareous clay soils and can tolerate drought and salt stress. Multivariate analysis of the vegetation of 100 sampled stands supporting growth of the three geophytic species in the study area led to the recognition of four vegetation groups namely, (A) Alhagi graecorum, (B) Cyperus capitatus, (C) Lycium schweinfurthii var. schweinfurthii-Asparagus stipularis and (D) Juncus acutus subsp. acutus. Vegetationally, the vegetation groups associated with the three species can be distinguished into two community types. The first one is psammophytic community comprising vegetation groups A and B that may represent the non-saline sand formations (flats, hummocks and dunes). The second one is halophytic community including vegetation groups C and D that may represent the saline sand flats and salt marsh habitat types, respectively. Sodium adsorption ratio, electrical conductivity, sodium cation, chlorides, silt and sand fractions, pH value, moisture content, bicarbonates and available phosphorus were the most effective soil factors that controlling the abundance and distribution of the plant communities associated with the investigated geophytes. This study showed the ecological features of the selected geophytes in terms of their habitats, associated plant communities and the most edaphic factors controlling their richness and distribution in the study area.

Micropropagation of Asparagus by in vitro shoot culture.

Asparagus officinalis is most extensively studied species within the genus Asparagus, which is well known as garden asparagus. This species is dioecious with unisexual flowers, which means that generative propagation gives roughly equal number of male and female plants. Male plants are high yielders and preferred commercially over female plants. Tissue culture techniques could efficiently promote vegetative propagation of male plants and pave the way for efficient plant breeding.This chapter describes an efficient micropropagation protocol for developing rapid growing in vitro Asparagus shoot cultures. The source of explants, inoculation, and shoot proliferation, followed by shoot propagation, rooting, and acclimatization is described. The optimal medium for Asparagus micropropagation described in this chapter is composed of MS macro- and microelements and a combination of auxins and cytokinins. Plant growth regulators NAA, kinetin, and BA were used in various concentrations. Three different media representing the whole micropropagation protocol of Asparagus are described; medium for shoot initiation, medium for shoot multiplication, and medium for root formation. By in vitro propagation of Asparagus, root initiation is difficult, but can be promoted by adding growth retardant ancymidol which also greatly promotes shoot development and suppresses callus formation.