High-frequency plant regeneration from leaf-disc cultures of Jatropha curcas L.: an important biodiesel plant

A simple, high-frequency and reproducible protocol for induction of adventitious shoot buds and plant regeneration from leaf-disc cultures of Jatropha curcas L. has been developed. Adventitious shoot buds were induced from very young leaf explants of in vitro germinated seedlings as well as mature field-grown plants cultured on Murashige and Skoog’s (MS) medium supplemented with thidiazuron (TDZ) (2.27 μM), 6-benzylaminopurine (BA) (2.22 μM) and indole-3-butyric acid (IBA) (0.49 μM). The presence of TDZ in the induction medium has greater influence on the induction of adventitious shoot buds, whereas BA in the absence of TDZ promoted callus induction rather than shoot buds. Induced shoot buds were multiplied and elongated into shoots following transfer to the MS medium supplemented with BA (4.44 μM), kinetin (Kn) (2.33 μM), indole-3-acetic acid (IAA) (1.43 μM), and gibberellic acid (GA₃) (0.72 μM). Well-developed shoots were rooted on MS medium supplemented with IBA (0.5 μM) after 30 days. Regenerated plants after 2 months of acclimatization were successfully transferred to the field without visible morphological variation. This protocol might find use in mass production of true-to-type plants and in production of transgenic plants through Agrobacterium/biolistic-mediated transformation.     

Jatropha curcas and assisted phytoremediation of a mine tailing with biochar and a mycorrhizal fungus

Soil pollution is an important ecological problem worldwide. Phytoremediation is an environmental-friendly option for reducing metal pollution. A greenhouse experiment was conducted to determine the growth and physiological response, metal uptake, and the phytostabilization potential of a nontoxic Jatropha curcas L. genotype when grown in multimetal-polluted conditions. Plants were established on a mine residue (MR) amended or not amended with corn biochar (B) and inoculated or not inoculated with the mycorrhizal fungus Acaulospora sp. (arbuscular mycorrhizal fungus, AMF). J. curcas was highly capable of growing in an MR and showed no phytotoxic symptoms. After J. curcas growth (105 days), B produced high desorption of Cd and Pb from the MR; however, no increases in metal shoot concentrations were observed. Therefore, Jatropha may be useful for phytostabilization of metals in mine tailings. The use of B is recommended because improved MR chemical properties conduced to plant growth (cation-exchange capacity, organic matter content, essential nutrients, electrical conductivity, water-holding capacity) and plant growth development (higher biomass, nutritional and physiological performance). Inoculation with an AMF did not improve any plant growth or physiological plant characteristic. Only higher Zn shoot concentration was observed, but it was not phytotoxic. Future studies of B use and its long-term effect on MR remediation should be conducted under field conditions.     

Global patterns of arthropod herbivory on an invasive plant,the physic nut (Jatropha curcas L.)

The physic nut (Jatropha curcas L.) is a multipurpose and oil‐producing shrub of Central and South American origin. Since the 15th century, this shrub has existed across tropical regions. Despite its presumed resistance to herbivores, reports show that arthropod herbivores infest it. However, no comprehensive account of arthropod herbivores, which consume the physic nut, exists. Here, we conducted a literature review that provides a comprehensive account of arthropod herbivores of the physic nut. Based on the co‐evolutionary hypothesis, we expected to find a higher herbivore of species richness and a larger proportion of native herbivores within the native range than elsewhere. As physic nut is a well‐defended plant chemically, we expected to find evidence for highest herbivory levels in plant parts that are the least defended. By the literatures review, we compiled 78 arthropod herbivores representing nine orders and from 31 families that feed on physic nut across the globe. As expected, the highest numbers of herbivores (34 species) were documented within the native range of the J. curcas and the lowest species number (21 species) in Africa. Of the 34 species in Central and South America, 94% were of native origin. Nine species were found feeding on J. curcas on more than one continent. Origins of 49% of species were from the native range of J. curcas. The highest percentage (54%) of species belonged to Hemiptera. With regard to feeding guilds, 59% of the herbivores belonged to sucking and 41% to chewing species. Forty‐one per cent of species were flower or fruit feeders, and 36% foliage feeders. We conclude that J. curcas is, despite its toxicity, vulnerable to herbivory, mainly to foliage, flower and fruit feeders.     

Morphogenesis and plant regeneration from cotyledonary cultures of Eucalyptus

Callus cultures were established from hypocotyls and cotyledons derived from young seedlings of Eucalyptus citriodora. Successful plantlet production from cotyledonary callus was achieved within 6 weeks on Murashige and Skoog's basal medium supplemented with zeatin (1 mg/l) and indoleacetic acid (0.2 mg/l). Leaf and shoot callus obtained from one-year-old plants did not differentiate. Results reported contribute to defining optimal conditions for callus growth and plantlet formation.     

Integrated genome sequence and linkage map of physic nut (Jatropha curcas L.), a biodiesel plant

The family Euphorbiaceae includes some of the most efficient biomass accumulators. Whole genome sequencing and the development of genetic maps of these species are important components in molecular breeding and genetic improvement. Here we report the draft genome of physic nut (Jatropha curcas L.), a biodiesel plant. The assembled genome has a total length of 320.5 Mbp and contains 27 172 putative protein‐coding genes. We established a linkage map containing 1208 markers and anchored the genome assembly (81.7%) to this map to produce 11 pseudochromosomes. After gene family clustering, 15 268 families were identified, of which 13 887 existed in the castor bean genome. Analysis of the genome highlighted specific expansion and contraction of a number of gene families during the evolution of this species, including the ribosome‐inactivating proteins and oil biosynthesis pathway enzymes. The genomic sequence and linkage map provide a valuable resource not only for fundamental and applied research on physic nut but also for evolutionary and comparative genomics analysis, particularly in the Euphorbiaceae.     

Evaluation of damage to physic nut (Jatropha curcas) by true bugs

The type of injury and amount of damage to the fruits and seeds of physic nut (Jatropha curcas L., Euphorbiaceae) caused by two species of fruit feeding true bugs, Pachycoris klugii Burmeister (Heteroptera: Scutelleridae) and Leptoglossus zonatus (Dallas) (Heteroptera: Coreidae), and the flower feeding true bug, Hypselonotus intermedius Distant (Heteroptera: Coreidae), were assessed using field cages. The parameters analysed were infructescence, fruit, seed and seed kernel weight, seed length, fruit and seed abortion, seed malformation and oil and protein content of the seed kernels. All three species reduced overall yield. The two members of the fruit feeding guild led to premature fruit abortion and malformed seeds. Reduction of fruit, seed and seed kernel weight as well as seed length could be expressed as linear regressions of adult population densities. The oil content of the seeds was slightly reduced by the bugs, but protein content remained unchanged. Damage increased with the developmental stage of the larvae. Whereas adult L. zonatus of both sexes produced more damage than the nymphs, female P. klugii caused less damage than nymphs, and male P. klugii no significant damage at all. H. intermedius is a pollinator of this entomogamous plant. At high densities, however, it reduced the number of fruits maturing.     

Somatic embryogenesis and plant regeneration from leaf, root and stem-derived callus cultures of Areca catechu

Plant regeneration through somatic embryogenesis of Areca catechu L. was established using leaf, root and stem segments as explants. Embryogenic callus was induced and maintained on medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) or 3,6-dichloro-2-methoxybenzoic acid (dicamba) at concentrations 2, 4, 6 and 8 mg dm⁻³ in darkness. Somatic embryos were found on primary callus in the presence of 2 and 4 mg dm⁻³ dicamba and during subculture on 2 – 8 mg dm⁻³ 2,4-D or 2 – 4 mg dm⁻³ dicamba-containing media. Plantlet conversion from embryos was successfully achieved on growth regulator-free medium. The plants grew well when transplanted to containers in shaded greenhouse.     

Factors influencing direct shoot regeneration from mature leaves of Jatropha curcas, an important biofuel plant

In order to establish a highly efficient and sustainable regeneration system, we systematically researched the key factors affecting direct shoot regeneration from Jatropha curcas leaves that were collected from Hainan (HN1-1), Lijiang (LJ3-1), and Yuxi (YX2-12) provinces in China. The L₉(3⁴) orthogonal test of thidiazuron (TDZ), kinetin (Kn), and gibberellic acid (GA₃) were studied, and the explant type, growth age, and cultivar of leaves were subsequently investigated. Simultaneously, the combinations of plant growth regulators (PGRs) promoting shoot bud proliferation, elongation, and root establishment were examined. The results showed that the best medium for shoot bud induction was Murashige and Skoog (MS) medium supplemented with 1.0 mg/L TDZ, 0.5 mg/L Kn, and 0.5 mg/L GA₃. TDZ was the key PGR, while Kn and GA₃ played an important role in shoot bud elongation and the number of shoots per leaf disk, respectively. The induced shoot buds proliferated and readily elongated in MS medium with 0.3 mg/L 6-benzylaminopurine and 0.01 mg/L indole-3-butyric acid (IBA) and established roots in half-strength MS medium supplemented with 2.0 mg/L IBA. Using the previously described methods, the third to fifth leaves were found to be the best explant source for shoot bud induction, with a high induction rate, large shoot numbers per disk, excellent proliferation, and consistent rooting. With the use of this regeneration system, the shoot bud induction rate increased from the reported rate of 53.5% to more than 90% using different explants and cultivars, and the shoot number per leaf disk (shoot length?≥?0.5 cm) increased from 1.6 to 3.5. Thus, this optimized regeneration system will effectively promote the propagation and genetic transformation of J. curcas.     

Long-duration,high-frequency plant regeneration from cereal tissue cultures

By visual examination of calli derived from germinating seeds of wheat, oats, rice, proso millet, and pearl millet it has been possible to visually select embryogenic (E) callus which, on transfer to a regeneration medium, forms plants an average of 33 times more frequently than non-embryogenic (NE) callus of equal mass. Embryogenic callus consists of small isodiametric cells averaging 31 m in diameter; NE callus consists of long tubular cells averaging 52 m in width and 355 m in length. Production of E callus is in many cases promoted by media containing 2,4-di- or 2,4,5-trichlorophenoxyacetic acid (2,4-D or 2,4,5-T) plus indole-3-acetic acid or tryptophan+kinetin. Production on NE callus is promoted by media containing 2,4-D or 2,4,5-T alone. As a result of initial experiments to optimize both media for E callus production and media for plant regeneration, callus derived in six passages from an average of 26 seeds could produce about 1,000 regenerated plants.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - 2,4,5-T 2,4,5-trichlorophenoxyacetic acid - IAA indole-3-acetic acid - Kin kinetin - Trp L-tryptophan - E embryogenic - NE non-embryogenic     

Plant regeneration through the direct induction of shoot buds from petiole explants of Jatropha curcas: a biofuel plant

An efficient and reproducible method for the regeneration of Jatropha curcas plants has been developed. The method employed direct induction of shoot buds from petiole explants, without the formation of an intervening callus using a Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ). The best induction of shoot buds (58.35%) and the number of shoot buds per explant (10.10) were observed when in vitro petiole explants were placed horizontally on MS medium supplemented with 2.27 µM TDZ after 6 weeks. The induced shoot buds were transferred to MS medium containing 10 µM kinetin (Kn), 4.5 µM 6-benzyl aminopurine (BAP) and 5.5 µM α-naphthaleneacetic acid (NAA) for shoot proliferation. The proliferated shoots could be elongated on MS medium supplemented with different concentrations and combinations of BAP, indole-3-acetic acid (IAA), NAA and indole-3-butyric acid (IBA). MS medium supplemented with 2.25 µM BAP and 8.5 µM IAA was found to be the best combination for shoot elongation and 3.01–3.91 cm elongation was achieved after 6 weeks. However, significant differences in plant regeneration and shoot elongation were observed among the genotypes studied. The orientation (horizontal or vertical) and source (in vitro or in vivo) of explants also significantly influenced plant regeneration. The elongated shoots could be rooted on half-strength MS medium supplemented with 2% sucrose, different concentrations and combinations of IBA, IAA and NAA, and 0.25 mg L⁻¹ activated charcoal. Half-strength MS medium supplemented with 2% sucrose, 15 µM IBA, 5.7 µM IAA, 5.5 µM NAA and 0.25 mg L⁻¹ activated charcoal was found to be the best for promoting rooting. The rooted plants could be established in soil with more than 90% survival.     

Rapid plant regeneration from various explants of Jatropha integerrima

A simple, rapid and reproducible protocol for direct shoot regeneration from different explants of Jatropha integerrima was developed. Prolific adventitious shoot bud initiation was obtained using a combination of 2.2 or 4.4 M benzyladenine and 4.9 M indole-3-butyric acid (IBA). Reduction of IBA concentration (2.5 M) promoted further development of shoots. Regenerated shoots rooted readily on Murashige and Skoog (MS) medium lacking growth regulators. Plantlets were acclimatized and successfully transferred to pots.     

Determination of antimicrobial and phytochemical compounds of Jatropha curcas plant

The aim of this study was to explore the effectiveness of different parts of J. curcas plant against some selected human pathogens as antimicrobial agent which are known to cause diseases and to check antioxidant and phytochemicals from different plant sections of J. curcas. Plant extracts were analyzed by quantification of antimicrobial and phytochemical compounds. This study reveals that 20% ethanol stem extract of J. curcas showed maximum antibacterial activity (40 ± 0.0 mm) against Klebsiella pneumonia. Water extract of root of J. curcas also inhibited E. coli (35.25 ± 0.35 mm). The growth of K. pneumonia and Agrobacterium tumifaciens were also ceased when ethanol extract of J. curcas root applied to check their potential as antimicrobial agent. The results also revealed that fungal species, Aspergillus niger, and Pencillium notatum noted the maximum antifungal activity in ethanol extract of flower and methanol extract of root (38.5 ± 0.7 mm) and (27.25 ± 0.35 mm) respectively. Phytochemicals and many secondary metabolites were present in J. curcas extracts such as alkaloids, steroids, tannins, glycosides, flavonoids, saponins, courmerin, and phenolic compounds. It also showed the highest density of color in the different parts of plant extract of J. curcas. Similarly, biochemical primary metabolites were observed at maximum amount of biochemical in different parts of J. curcas, and correlated with antimicrobial activity. The study concluded that J. curcas has great potential as antibacterial agent and cure various human pathogens.     

木本油料植物麻疯树的研究进展

近来,麻疯树(Jatropha curcas L.)因可以作为生物柴油的原料而受到广泛关注。虽然麻疯树具有含油量高且可在边际土地种植的优势,但同时存在理想品种缺乏、种植面积限制、基础研究薄弱等方面的问题。为了使常规育种和生物技术改良工作者更好地了解麻疯树的研究进展,确定日后麻疯树育种和遗传改良的方向,综述了麻疯树种质资源与遗传多样性分析、生理及对环境适应性、组学研究、油脂合成功能基因研究、育种等方面国内外的最新进展。     

Identification and host range of causal agent of dieback disease on Jatropha curcas

Diseased Jatropha branches collected from Institute for Agricultural Research (IAR), Samaru, Zaria field were plated in Potato Dextrose Agar with Streptomycin (PDAS) and incubated for seven days. Grown cultures were observed under microscope and with aid of identification manuals Fusarium sp. was isolated. Inoculated seedlings showed symptoms of dieback. The fungus was also inoculated on some crop plants and it induced the typical dieback symptoms on sesame and leaf spot/ blight on groundnut and soya beans, respectively.     

Plant regeneration of non-toxic Jatropha curcas—impacts of plant growth regulators, source and type of explants

Jatropha curcas is an oil bearing species with multiple uses and considerable economic potential as a biofuel plant, however, oil and deoiled cake are toxic. A non-toxic variety of J. curcas is reported from Mexico. The present investigation explores the effects of different plant growth regulators (PGRs) viz. 6-benzyl aminopurine (BAP) or thidiazuron (TDZ) individually and in combination with indole-3-butyric acid (IBA), on regeneration from in vitro and field-grown mature leaf explants, in vitro and glasshouse-grown seedlings cotyledonary leaf explants of non-toxic J. curcas. In all the tested parameters maximum regeneration efficiency (81.07%) and the number of shoot buds per explants (20.17) was observed on 9.08 μM TDZ containing Murashige and Skoog’s (MS) medium from in vitro cotyledonary leaf explants. The regenerated shoot buds were transferred to MS medium containing 10 μM kinetin (Kn), 4.5 μM BAP and 5.5 μM α-naphthaleneacetic acid (NAA) for shoot proliferation. The proliferated shoots could be elongated on MS medium supplemented with 2.25 μM BAP and 8.5 μM IAA. Rooting was achieved when the basal cut end of elongated shoots were dipped in half strength MS liquid medium containing different concentrations and combinations of IBA, IAA and NAA for four days followed by transfer to growth regulators free half strength MS medium supplemented 0.25 mg/l activated charcoal. The rooted plants could be established in soil with more than 90% survival rate.     

Chemical Constituents from Roots of Jatropha curcas

Thirteen compounds were isolated from the roots of datropha curcas L. Combining the determination of physico-chemical constants and spectral analyses (IR, 1H-NMR, 13C-NMR, EIMS, FABMS), the structures of the compounds were identified as 5α-stigmastane-3, 6-dione (1), nobiletin (2), β-sitosterol (3), taraxerol (4), 2S-tetracosanoic acid glyceride-1(5),5-hydroxy-6,7-dimethoxycoumarin (6), jatropholone A (7), jatropholone B (8), 6-methoxy-7-hydroxycoumarin (9), caniojane (10), 3-hydroxy-4-methoxybenzaldehyde (11), 3-methoxy-4-hydroxybenzoic acid (12) and daucosterol (13). Among them, compound 5 is a new compound which has never been reported in China and abroad, compound 1, 2, 9, 10, 11, 12 were first time isolated from the plant, 7 and 8 are a pair of stereoisomers which can be inverted in dilute basic solution. 10 is a diterpenoid containing peroxide bridge.     

Morphogenesis and plant regeneration from cultured endosperm of Emblica officinalis Gaertn

Mature endosperm of Emblica Officinalis (Euphorbiaceae) formed a continously growing callus on MS medium supplemented with an auxin (2,4-D or IAA) and a cytokinin (K or BAP). Subculture of callus on MS with BAP (0.2 mg/l) and IAA (0.1 mg/l) resulted in formation of shoots and embryo-like structures in 50 and 8 per cent cultures, respectively. Regeneration of shoots was more frequent when both BAP (0.2 mg/l) and IAA (0.1 mg/l) were present than on BAP (0.2 mg/l) alone. The embryo-like structures produced plantlets.Abbreviations BAP benzylaminopurine - 2,4-D 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - IAA indoleacetic acid - K kinetin - NAA naphthaleneacetic acid - PDB para-dichlorobenzene (née Arora)