Lipids are key components of all living cells. Acyl lipids and sterols provide the matrix of the biological membranes that both define the boundaries of cells and organelles, and act as sites for the trafficking of molecules within and into/out of cells. Lipids are also important metabolic intermediates and the most efficient form of energy storage that is available to a cell. It is the latter, energy-storing function that is of most relevance to this review. Storage lipids are accumulated in abundance in many of our most important crops, including maize, soybean, rapeseed, and oil palm, giving rise to a commercial sector valued at over $50 billion/year. Because the storage lipids of the major global oil crops have a relatively restricted composition, there is great interest in using all available breeding technologies, whether traditional or modern, to enhance the variation in lipid quality in existing crops and/or to domesticate new crops that already accumulate useful novel lipids. Over the past few decades, there has been a great deal of effort to manipulate fatty acid composition in order to produce novel lipids, especially for industrial applications. However, these attempts, many based on genetic engineering, have met with only limited commercial success to date. More recently, there has been a resurgence of interest in the modification of both acyl and non-acyl lipids to enhance the nutritional quality of plant oils. In this review, we will examine the background to plant lipid modification and some of the latest developments, with a particular focus on edible oils.

Proteinase inhibitor II (PIN2) proteins from the Solanaceae family have been previously used in plant transformation to acquire protection against caterpillars. Some of these PIN2 proteins have been shown to exhibit exogenous activities against trypsin and/or chymotrypsin in vitro. Despite their application in conferring insect resistance in transgenic plants, the endogenous roles of this family of proteins in various plant species have not been well defined. To investigate the exogenous and endogenous functions of PIN2 proteins, cDNAs encoding PIN2 proteins from the weed Solanum americanum (American black nightshade), designated SaPIN2a and SaPIN2b, were cloned and characterized. The localization of S. americanum SaPIN2a and SaPIN2b mRNAs and proteins in the reproductive tissues destined to undergo developmental programmed cell death subsequently led to investigations into their function during seed development. Using plant transformation of lettuce and S. americanum, it was evident that: (1) the expression of SaPIN2a in transgenic lettuce conferred resistance to cabbage looper (Trichoplusia ni) caterpillars; and (2) the expression of siRNAs from a PIN2–RNAi construct resulted in transgenic S. americanum that were impaired in seed development. These results suggest that S. americanum PIN2 proteins not only enhance resistance to caterpillars (when expressed exogenously) but they function in inhibiting endogenous proteases that are expressed during seed development. Specifically, the aborted seeds of PIN2–RNAi lines showed abnormal endothelium that subsequently affected endosperm and embryo development.

A method was developed for plant regeneration from alginate-encapsulated shoot tips of Phyllanthus amarus. Shoot tips excised from in vitro proliferated shoots were encapsulated in calcium alginate beads. The best gel complexation was achieved using 3% sodium alginate and 75 mM CaCl₂·2H₂O. Maximum percentage response for conversion of encapsulated shoot tips into plantlets was 90% after 5 wk of culture on Murashige and Skoog (MS) medium without plant growth regulator. The regrowth ability of encapsulated shoot tips was affected by the concentration of sodium alginate, storage duration, and the presence or absence of MS nutrients in calcium alginate beads. Plantlets with well-developed shoot and roots were transferred to pots containing an autoclaved mixture of soilrite and peat moss (1:1). The conversion of encapsulated shoot tips into plantlets also occurred when calcium alginate beads were directly sown in autoclaved soilrite moistened with 1/4-MS salts. Encapsulation of vegetative propagules in calcium alginate beads can be used as an alternative to synthetic seeds derived from somatic embryos.

We report a protocol for efficient plant regeneration of four tall fescue (Festuca arundinacea Schreb.) cultivars (‘Surpro’, ‘Coronado’, ‘Summer Lawn,’ and ‘Fawn’) via somatic embryogenesis. Calli were initiated from mature seeds grown on modified Murashige and Skoog (MMS) medium supplemented with 7.0 mg l⁻¹ (31.7 μM) 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.05 mg l⁻¹ (0.23 μM) kinetin (Kin). Calli were maintained and proliferated by subculture at monthly intervals on MMS medium containing 4.5 mg l⁻¹ (20.4 μM) 2,4-D and 0.2 mg l⁻¹ (0.9 μM) Kin. Somatic embryos (SE) were induced from seed-derived calli on SE-induction medium (MMS supplemented with 2.0 mg l⁻¹ 2,4-D and 0.2 mg l⁻¹ Kin). Plantlets were regenerated from somatic embryogenic calli grown on modified SH medium supplemented with 2 mg l⁻¹ Kin. Using this optimized protocol, 78.6–82.3% of mature seeds of all four cultivars produced SE clusters, of which 93.5–95.3% regenerated into plants within 10 wk. The regenerants showed no phenotypic abnormalities.

Plant regeneration through direct somatic embryogenesis was achieved from root segments derived from in vitro shoots of Rauvolfia micrantha Hook. f. (Apocynaceae) grown for 6 wk in half-strength Murashige and Skoog (MS) medium with 3% sucrose, 100 mg l⁻¹ myo-inositol, and 0.5 mg l⁻¹ α-naphthaleneacetic acid (NAA). The effects of photoperiod and plant growth regulators (PGRs) in half-strength MS medium were studied for the rapid and maximum induction of somatic embryos. The characteristic globular or heart-shaped stages of somatic embryogenesis were not found and cotyledonary stage embryos occasionally appeared without the intervention of callus in total darkness and 16-h photoperiod. Root segments cultured in the medium containing 0.1 mg l⁻¹ NAA and 0.2 mg l⁻¹ 6-benzyladenine (BA) under 16-h photoperiod showed the maximum frequency (39%) of embryogenesis. The frequency of embryo formation was increased to 63% when they were cultured in medium with 0.1 mg l⁻¹ NAA and 0.2 mg l⁻¹ BA in the dark for 4 wk, then grown under the 16-h photoperiod. Explants with developing embryos developed into plants after transfer to half-strength MS medium supplemented with 0.1 mg l⁻¹ BA and 0.05 mg l⁻¹ NAA. The well-developed plants were hardened and most plants (80%) survived and were phenotypically similar to the mother plants.

A protocol for the induction of somatic embryogenesis from immature zygotic embryos of Rosa bourboniana, a scented rose species, was established. Somatic embryos were induced after 8 wk of inoculation of zygotic embryos on MS medium supplemented with different concentrations of 2,4-dichlorophenoxy acetic acid (5–15 μM). In addition to 2,4-dichlorophenoxy acetic acid concentrations, somatic embryogenesis was also influenced by the month of collection of the explant and the stage of maturity of the hip. Maximum embryogenic response (16.6%) was observed using 2,4-dichlorophenoxy acetic acid (15 μM), from green hips in the month of September. The use of l-proline (800 mg l⁻¹) was found to be optimum for secondary embryogenesis. On periodic subculturing, the cultures formed somatic embryos sustainably over a period of 18 mo. For somatic embryo germination, 6-benzylaminopurine (5 μM) was found to be most suitable. Rooted plants were transferred successfully to soil and appear morphologically normal under greenhouse conditions. Transfer of plants for hardening was most suitable during the active growth period between June and September.

This study reports an improved protocol for in vitro-shoot multiplication and ex vitro acclimation of Bupleurum kaoi, an endangered medicinal herb. Nodal segments were cultured in half-strength Murashige and Skoog (MS) basal medium supplemented with different concentrations of benzyladenine (BA) and kinetin. The presence of 0.25 mg l⁻¹ BA induced the highest number of shoots per explant after 8 wk of culture. Although BA was more effective than kinetin on shoot multiplication, it induced hyperhydric shoots at all concentrations tested. The use of dispense paper (DP) instead of aluminum foil (AF) for container closure was found to reduce hyperhydricity and improve ex vitro acclimation. The best survival rate (61%) was obtained when plantlets were grown in MS basal medium containing 0.5 mg l⁻¹ indole-3-butyric acid and 0.1–0.2 mg l⁻¹ α-naphthaleneacetic acid using DP as container closure. Leaves of the plant treated with AF6 (two layers of AF as container closure and 6 wk of incubation) lacked epicuticular wax and possessed larger stomata, higher stomata density, and fewer functional stomata compared to those of plants treated with AF2 DP4 (two layers of AF for 2 wk, then replaced AF by three layers of DP for 4 wk) and ex vitro-acclimated plantlets.

This study reports a protocol for plant regeneration from cultured explants of green gram [Vigna radiata (L.) Wilczek] via somatic embryogenesis. Somatic embryos were induced from mature cotyledons of var. TAP-7 and Pusa Baisaki when cultured on Murashige and Skoog (MS) medium fortified with different concentrations of 2,4-dichlorophenoxyacetic acid, α-naphthaleneacetic acid (NAA) and 2,4,5-trichlorophenoxyacetic acid singly or in combination with 2.22–8.88 μM N⁶-benzylaminopurine (BAP) or 2.32–9.38 μM kinetin. The type and concentration of auxin and plant genotype influenced the frequency of somatic embryogenesis. NAA was the most effective auxin for somatic embryo induction. The well-developed, cotyledonary shaped embryos of var. TAP-7 germinated into plantlets at a frequency of 56.6% on MS medium supplemented with 1.88 μM abscisic acid and 6.66 μM BAP. Regenerated plants were transferred to soil and grown to maturity with 90% survival. The protocol described here offers a good potential for genetic improvement using gene transfer techniques and the production of synthetic seeds of V. radiata.

Micropropagation via enhanced axillary shoot proliferation was investigated in the ornamental Eucalyptus cv. ‘Urrbrae Gem’ using in vitro germinated seedlings and was successfully achieved using woody plant medium (WPM) supplemented with 2.2 μM benzylaminopurine, 1.0 μM α-naphthaleneacetic acid, and 1.5 μM gibberellic acid (GA₃), gelled with 5 g l⁻¹ Phytagel^®. Shoot proliferation was greater on WPM and QL media with GA₃ compared to B5, AP, and TK media with or without GA₃. GA₃ was required for shoot elongation as the internodes were otherwise very short and unsuitable for multiplication or root initiation. Root initiation was improved using (1/2)WPM supplemented with 20 μM indole-3-butyric acid (IBA) over a 7 d pulse, followed by subculture to IBA-free medium, compared to placing shoots on low levels of IBA for 4–6 wk. Plantlets were successfully hardened off to the natural environment via a fogger at 67% relative humidity at 21°C for 3 d and continued to thrive as potted plants. This is the first report of successful micropropagation in an ornamental eucalypt (subgenus Symphyomyrtus) from seedling explants.

This study reports a protocol for high-efficiency plant regeneration from leaf explants of male Himalayan poplar (Populus ciliata Wall.). Shoots were regenerated at high frequencies from explants grown on Murashige and Skoog (MS) medium supplemented with 0.5 mg l⁻¹ kinetin and 0.2 mg l⁻¹ indole-3-acetic acid (IAA). Regenerated shoots developed roots in MS medium supplemented with 0.1 mg l⁻¹ IAA. Himalayan poplar plantlets could be produced within 2 mo. after acclimatization in a sterile mixture of sand and soil.

A method was developed for in vitro regeneration of plants via somatic embryogenesis in Chorispora bungeana, an alpine plant with freeze-tolerance, using cell suspensions initiated from leaf-derived callus. Primary calli were induced from leaves of C. bungeana grown on Murashige and Skoog (MS) media supplemented with 4.0 mg l⁻¹ gibberellic acid (GA₃), 0.2 mg l⁻¹ α-naphthaleneacetic acid (NAA) and 0.2 mg l⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D). Suspension culture was initiated by incubating the callus particulates in liquid MS medium supplemented with 1.0 mg l⁻¹ kinetin (KT) and 0.2 mg l⁻¹ NAA. Individual early cotyledonary-stage somatic embryos isolated from cell suspension developed into whole plants on medium containing high levels of sucrose (60 and 90 g l⁻¹), whereas lower sucrose concentrations (0 and 30 g l⁻¹) were inhibitory to main root development. On the MS medium with 90 g l⁻¹ sucrose, one regenerated plant exhibited hetero-morphologic leaves, while other plants grown on different media showed a transformation from stem to root.

The present investigation reports optimized parameters for somatic embryogenesis and cryopreservation of embryogenic cultures using shoot apical domes from mature trees of Pinus roxburghii Sarg. Embryogenic tissue of P. roxburghii Sarg. was cryopreserved for 24 h, 10 d, and 8 wk using sorbitol and dimethylsulfoxide (DMSO) as cryoprotectants. Results indicate that 0.2 M sorbitol and 5% DMSO had the best cryoprotecting effect. The recovered tissue showed luxuriant growth on maintenance medium (II). Partial desiccation of thawed embryogenic tissue for 24 h prior to transfer to maturation medium enhanced the maturation of somatic embryos. Maturation frequency increased from 1.3 to 18.3% after 12 h desiccation treatment, and from 18.3 to 61.8% after 24 h of desiccation. However, non-desiccated embryogenic tissue produced the least number of somatic embryos (1.3%) on the maturation medium with the same abscisic acid and Gellan gum concentration. All the three embryogenic lines produced plantlets and had the same appearance and normal growth as compared to unfrozen controls.

The in vitro culture responses from different explants of a race-specific resistant cultivar (Red Mexican) and a race-susceptible cultivar (Palmeña) to halo-blight pathogen (Pseudomonas syringae pv. phaseolicola) were studied. Two kinds of filtrate obtained from a phaseolotoxin producer wild type and a non-producer mutant of P. syringae pv. phaseolicola race-7 were used. Callus formation of Red Mexican was significantly reduced in the presence of phaseolotoxin. Bud-shoot growth was more sensitive than callus formation to other metabolites present in the pathogen filtrate, but the presence of phaseolotoxin in the media showed a positive correlation between resistance to halo blight race-7 pathogen and bud-shoot growth. Our results indicate that differential in vitro responses are influenced by the plant genotype and by the metabolite composition and concentration of the filtrate.

Conventional breeding methods have not been very successful in producing pest-resistant genotypes of pigeonpea, due to the limited genetic variation in cultivated germplasm. We have developed an efficient method to produce transgenic plants of pigeonpea by incorporating the cry1Ab gene of Bacillus thuringiensis through Agrobacterium tumefaciens-mediated genetic transformation. The novel tissue culture protocol is based on the direct regeneration of adventitious shoot buds in the axillary bud region of in vitro germinating seedlings by suppressing the axillary and primary shoot buds on a medium containing a high concentration of N⁶-benzyladenine (22.0 μM). The tissue with potential to produce adventitious shoot buds can be explanted and used for co-cultivation with A. tumefaciens carrying the synthetic cry1Ab on a binary vector and driven by a CaMV 35S promoter. Following this protocol, over 75 independently transformed transgenic events of pigeonpea were produced and advanced to T2 generation. Amongst the recovered primary putative transformation events, 60% showed positive gene integration based on initial polymerase chain reaction (PCR) screening. PCR analysis of the progenies from independent transformants followed gene inheritance in a Mendelian ratio and 65% of the transformants showed the presence of single-copy inserts of the introduced genes. Reverse transcription–polymerase chain reaction analysis showed that the transcripts of the introduced genes were normally transcribed and resulted in the expression of Cry1Ab protein in the tested T2 generation plants. Interestingly, the content of Cry1Ab protein as a percent of total soluble protein varied in different tissues of the whole plant, showing the highest expression in flowers (0.1%) and least in the leaves (0.025%) as estimated by enzyme-linked immunosorbent assay. The transgenic plants produced in this study offer immense potential for the improvement of this important legume of the semi-arid tropics for resistance to insect pests.

This study reports the characterization of 11 plants regenerated from electrically fused protoplasts between Populus alba and Alnus firma. Growth characteristics of five regenerated plants (AP-1–AP-5) in terms of shoot height and leaf color showed small differences compared with those of P. alba grown in pots, and showed no difference in shoot height and diameter compared with those grown in nursery field. There was also no difference in the RAPD pattern between the plants regenerated from interfamilial protoplast fusion and P. alba. In contrast, the lately regenerated plants (AP-6–AP-11) grown in pots showed a marked difference in leaf morphology and RAPD pattern. There was a variation in the ratio of longitudinal to transverse length of leaves among the 11 plants from interfamilial fusions compared with that of protoclones and intraspecific fused protoplasts of P. alba.

This study describes the development of electrofusion techniques using the ‘donor–recipient’ model for the production of cybrids between common cultivated winter wheat (Triticum aestivum L.) cv. Jinghua No. 1 and a phylogenetically remote, sexually incompatible grass species, Italian ryegrass (Lolium multiflorum Lam.), which belong to two different subtribes: Triticinae and Loliinae. Wheat protoplasts were metabolically inactivated by iodoacetamide before fusion, while protoplasts of Italian ryegrass were X-ray irradiated before protoplast isolation. The suspension cells were directly used to optimize the inactivation parameters. By exploring the minimum irreversible membrane breakdown strength, the electrofusion parameters were optimized just a few minutes before electrofusion began. A total of 108 green plantlets were obtained, and about half of the green plants uncontrollably necrotized. Among all green plants, 14 were rooted normally and transplanted in growth chamber or field and developed to maturity. All these transplanted plants were male sterile with smaller and off-white anthers. Seeds were obtained by crossing with Jinghua No. 1. Three transplanted regenerants possessed the characteristics of glume facing the rachis, which was the taxonomic characteristic distinguishing the two subtribes of Triticinae and Loliinae. Although Southern blot hybridization analysis of 33 randomly selected regenerants using a wheat ribosomal DNA probe (pHA71) did not find any differences to wheat, analysis using two mitochondrial probes B342 (cox I), 490 (Pro II) and one chloroplastidic probe pHvc P5 revealed that 31 plants were ‘true cybrids’ by showing ryegrass-specific band(s) or new band(s). It also showed that the mitochondria and chloroplasts were not coexistent as the restriction fragment length polymorphism band of Italian ryegrass was not detected by the mitochondrial probes 7 (26s), B342 (cox I), pHJ2-7-1 (cox II), B30 (atp9), and the chloroplast probe pHvc P5. To regenerate the cybrids, the regeneration capacity of the recipient (wheat) was crucial in this study.

A morphological variant obtained from in vitro corm-derived plants of banana (Musa sp.) cv. Grande Naine (AAA) was evaluated up to harvest, and the genetic basis of variation was confirmed by the random amplified polymorphic DNA (RAPD). The corms formed during the multiplication phase of shoot tip-derived cultures of the cv. Grande Naine grown on Murashige and Skoog (MS) medium enriched with 13.3 μM N⁶-benzyladenine (BA) developed numerous morphological variants after transfer to MS medium with 6.66 μM BA. The variant designated as CUDBT-B1, with distinct morphological features, was further evaluated. The morphological features of CUDBT-B1 were variegated leaf, pseudostem, bracts, ovary of the male flower and fruits, reduced height, decreased lamina length and breadth, and early flowering. These features were also manifested in the second-cycle progeny of CUDBT-B1. RAPD assay showed a marker DNA band of 1650 bp, and differential band intensity between the CUDBT-B1 and normal clone. CUDBT-B1 was multiplied using shoot tip culture, and the shoots were rooted on half-strength MS medium supplemented with 2.69 μM α-naphthaleneacetic acid. All plantlets showed variegated leaves under field conditions.

A micropropagation protocol for Pouteria lucuma R. and Pav. var. La Molina was developed. Shoots from zygotic embryos with a portion of endosperm were established in vitro on Murashige and Skoog (MS) medium with 0.47 μM kinetin (Kin) and 0.54 μM naphthaleneacetic acid (NAA). Multiplication of shoots was accomplished using subapical shoots. The best axillary-shoot production was observed on MS basal medium with 2.2 μM benzyladenine (BA), 0.5 μM NAA, 1.4 μM gibberellic acid (GA₃), and 40 mg l⁻¹ adenine sulfate, with the development of up to three axillary shoots per subapical shoot. One hundred percent rooting was obtained from shoots grown for 4 wk on MS medium with 246 μM indole-3-butyric acid under light conditions. Eighty percent of the microplantlets survived after acclimatization when transplanted to a substrate previously enriched with beneficial soil bacteria. This study describes, for the first time, arbuscular mycorrhizal (AM) colonization of this species. Inoculation with AM fungi improved growth and development of lucumo plants and induced changes to the root morphology.

Rooting in vitro was examined for lentil nodal segments to test a recently published conclusion that shoot orientation has an effect on rooting. As is the case for many species, roots initiated and grew only at the proximal end of the microcutting regardless of its orientation. When the proximal end was in agar (a hypoxic environment) the rooting percentage was low (9–25%) even when the orientation of the microcutting was altered by inverting the culture tube. In contrast, when the proximal end of the microcutting was in an aerobic environment (from the shoot being placed upside down in agar medium or placed normally or upside down in an aerated medium), rooting percentages were higher (62–100%). Medium aeration at the proximal end of the microcutting is more important than shoot orientation for in vitro rooting of lentil microcuttings.