Volume 7 Number 1 2013

CONTENTS AND ABSTRACTS

Jaime A. Teixeira da Silva (Japan) Orchids: Advances in Tissue Culture, Genetics, Phytochemistry and Transgenic Biotechnology (pp 1-52)

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Review: Orchids include some of the world’s most important floricultural (cut-flower) and ornamental (pot and garden) plants, some of which have pharmacological interest. Even though seed propagation has been recorded since the early 19^th century, their micropropagation and tissue culture dominated orchid biotechnology until the 1990’s, while transformation and molecular technologies are now being more intensely focused. This review highlights literature until about 2005 that pertains to ornamental orchid in vitro cell, tissue and organ culture, micropropagation, genetics and transformation, and takes an in depth analysis at how each of these disciplines has influenced the use of biotechnology in the improvement and preservation of orchids around the world allowing orchid research to take a new direction in recent years. Closely related to this is the use of phytochemicals and secondary metabolites from orchids, which serve important medical and industrial purposes. Using biotechnology, these phytochemicals can be further explored and manipulated in vitro.

Debashish Behera, Chandi C. Rath, Umaballava Mohapatra (India) Medicinal Orchids in India and their Conservation: A Review (pp 53-59)

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Mini-Review: Application of traditional knowledge for the utilization of natural products, particularly of plant origin, has gained importance in the past several decades. For the tribal people of different parts of India, there is limited alternative to herbal medicines, which they have used for time immemorial. Along with other medicinal plants, orchids are considered to be an important source of herbal medicine. Orchids are among the most diverse of the flowering plant families, with over 181 genera and 1229 species specific to India. Orchids, which are well known for their floriculture value, are also used for curing several diseases. Due to over-exploitation for medicinal use and for the cut-flower trade, many orchids have become either rare or endangered. This review attempts to summarize the use of micropropagation to conserve Indian orchids of medicinal significance.

David C. Zlesak, Kathy Zuzek, Stan C. Hokanson (USA) Gender Inheritance and Identification of Male Sterility Gene RSMS1 in Intra- and Inter-specific Crosses of Dioecious Rosa setigera Michaux (pp 60-64)

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Original Research Paper: Rosa setigera Michaux (prairie or blackberry rose; 2n=2x=14) is the only dioecious rose species and only member of the Synstylae section (=Systylae) native to North America. Although flowers have male and female structures to attract pollinators, only one gender is typically functional per genotype. Intra- and interspecific crosses were made to document gender segregation in progeny. Seventeen of the 19 intraspecific crosses did not deviate from a 1:1 female:male ratio. The remaining two families shared a parent, with one family having significantly more and the other less of each gender. Interspecific crosses were only successful with female R. setigera genotypes, indicating the existence of unilateral interspecific cross incompatibility. All F₁ hybrids were male-sterile and female-fertile. Segregation for male fertility was observed in subsequent generations. Segregation data support the conclusion that a single gene (RSMS1) controls male sterility with a dominant allele needed for the male-sterile phenotype. This gene has high penetrance, but in some interspecific populations has weakened expressivity as seen by very low rates of in vitro pollen germination (<1%) and abnormal pollen tube growth. Potential applications and new research opportunities related to these advancements in understanding interspecific cross compatibility and gender inheritance in R. setigera are discussed.

Saranjeet Kaur, K. K. Bhutani (India) In Vitro Propagation of Paphiopedilum spicerianum (Reichb. F.) Pfitz. (pp 65-70)

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Original Research Paper: The goal of this study was to investigate various factors affecting germination in vitro and to establish a protocol for propagation of Paphiopedilum spicerianum which would provide information on various events of seed germination and seedling development thus helping to propagate and conserve this species. Capsules were harvested at two different stages of development. Six asymbiotic orchid seed germination media (Knudson C (KC), modified Knudson C (MKC), Terrestrial orchid medium (BM), modified Terrestrial orchid medium (BM1), Vacin and Went (VW) and Nitsch and Nitcsh (N)) were examined for their effectiveness in promoting seed germination, protocorm and seedling development of P. spicerianum. Besides checking the efficacy of different media, the effect of capsule maturity level, photoperiod (0/24 h; 12/12 h L/D), activated charcoal, plant growth regulators [6-benzyladenine (BA) and α-naphthalene acetic acid (NAA)] on seed germination was also assessed. The seeds from undehisced green capsules germinated with better frequency than seeds from mature burst capsules. Germination occurred regardless of media type. Amongst all media tested, highest germination percentage (62.75 ± 2.63%) was achieved in BM1 and activated charcoal (AC) under continuous darkness. The addition of NAA (1.5 mg l^-1) to medium resulted in the early formation of seedlings within 21.05 ± 0.05 weeks. Higher concentration of BA reduced the percentage of seed germination. After germination for 8 weeks in total darkness at the protocorm stage, a shift from darkness to light conditions (12/12 h L/D) was required for differentiation of protocorms into seedlings. Chlorophyll development was a post-protocorm phenomenon in the cultures. The current study has the potential to assist with the future development of ex situ conservation of this endangered species by producing innumerable viable seedlings.

Kathryn Kamo (USA) Optimized Conditions for Biolistic-mediated Transformation of Lilium longiflorum ‘Nellie White’ (pp 71-76)

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Original Research Paper: A variety of tissues were used for biolistic-mediated transformation of Lilium longiflorum ‘Nellie White’. Transgenic plants were not recovered from five-month-old, “non-select” callus or suspension cells that had been bombarded with pDM327 that contains the bar-uidA fusion gene under control of the CaMV 35S promoter. In comparison, ten transgenic plants were recovered from “select” callus that had been selected for its embryogenic-like appearance. Transgenic plants were also obtained following direct bombardment of precultured bulb scales. Both preculture time and preculture medium had an impact on transformation frequencies. Bulb scales cultured overnight on Murashige and Skoog’s medium containing 1.0 mg/L picloram before bombardment with 0.6 µm gold resulted in a transformation frequency of 0.9%. A one week or one month preculture time on Murashige and Skoog’s medium containing 1.0 mg/L picloram yielded higher, 4.2% and 5%, transformation frequencies, respectively. When bulb scales were cultured on Murashige and Skoog’s medium with either 2.0 mg/L dicamba or 0.5 mg/L picloram, lower frequencies of transformation, 1.2% and 0.8%, respectively, were observed. Direct bombardment of bulb scales eliminates the time needed for callus induction and multiplication when embryogenic callus is bombarded.

Jaime A. Teixeira da Silva (Japan), Apiradee Uthairatanakij, Kullanart Obsuwan (Thailand), Kazuhiko Shimasaki, Michio Tanaka (Japan) Elicitors (Chitosan and Hyaluronic Acid) Affect Protocorm-Like Body Formation in Hybrid Cymbidium (pp 77-81)

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Original Research Paper: Protocorm-like body (PLB) production from hybrid Cymbidium Twilight Moon ‘Day Light’ was examined on a new medium, Teixeira Cymbidium (TC) medium in response to chitosan and other elicitors. TC_PLB, the control medium specific for induction of PLBs, contained a unique set of micro- and macro-nutrients, 0.1 mg/l α-naphthaleneacetic acid and 0.1 mg/l kinetin, 2 g/l tryptone and 20 g/l sucrose, and solidified with 8 g/l Bacto agar. Crude deacetylated chitin (i.e., chitosan), chitosan and hyaluronic acid (HA) had a pronounced effect on PLB formation, depending on the concentration, when added to TC medium without plant growth regulators (PGRs). In all cases, the performance was never better than TC, although 0.1-1.0 mg/l chitosan or HA produced more PLBs/PLB and a greater fresh weight than TC basal medium without PGRs. This indicates that both these elicitors could induce PLBs and thus acted as phytohormones. Peroxidase activity, a signal of stress induction, increased in PLB extracts of PLBs grown on medium containing chitosan and HA, peaking when these elicitors were supplied at 50 mg/l.

Babita Saha, Animesh K. Datta, Siraj Datta (India), Jaime A. Teixeira da Silva (Japan) In Vitro Corm Development, Field Evaluation and Determination of Genetic Stability of Corm-derived Elite Gladiolus Germplasm(pp 82-85)

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Original Research Paper: An efficient protocol for in vitro corm production of Gladiolus (cv. ‘Green Bay’, ‘Intrepid’, ‘Sabnam’, ‘White Friendship’ and ‘Red Ginger’) was developed using liquid culture and coir as a matrix. The initial culture was established from the basal portions of the innermost leaves in Murashige and Skoog (MS) solid basal medium with 2.0 mg l^-1 α-naphthaleneacetic acid (NAA). Adventitious shoot buds developed (MS supplemented with 0.2 mg l^-1 NAA and 2.0 mg l^-1 6-benzyladenine) from responding callus (10-15 per callus culture) and elongated in solid MS with 0.5 mg l^-1 NAA. High regeneration frequency of corms (98.90%) was established when individual plantlets were cultured in MS liquid medium supplemented with 6% sucrose and 0.5 mg l^-1 NAA. In vitro corms were successfully cultivated in the field (‘White Friendship’ – high altitude cultivar; and ‘Sabnam’ – plain land cultivar) for two consecutive seasons and flowering was noted in the second generation of plants. Using RAPD profiles, the genetic fidelity of randomly selected in vitro and ex vitro corms of two cultivars (‘White Friendship’ and ‘Sabnam’) as well as that of their mother corms was compared.

Iftikhar Ahmad (Pakistan/USA), Muhammad Hussnain Shah (Pakistan) Trichoderma and Intercropping Impact Production, Quality and Corm Rot Disease of Gladiolus grandiflorus (pp 86-93)

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Original Research Paper: The effects of Trichoderma harzianum Rifai application and intercropping with Gazania rigens L. (treasure flower) or Calendula officinalis L. (pot marigold), members of the Asteraceae family, were studied on growth, yield, quality and controlling corm rot disease of gladiolus. Two experiments were conducted; first using healthy corms of two gladiolus cultivars, ‘Fado’ and ‘Cantate’ and second on both healthy and artificially infected (with Fusarium oxysporum) corms of cultivar ‘Fado’ in order to compare the individual treatment effects. The addition of Trichoderma to the medium increased growth, production and quality as well as reduced disease incidence when applied to healthy corms, although intercropping had a similar effect. When individual treatment effects were evaluated, use of both Trichoderma and intercrops, applied to infected corms, significantly improved plant height (65%), foliage growth (122%), reduced crop harvest time (23%), and disease (corm rot) incidence (78%), increased leaf area (70%), total leaf chlorophyll contents (162%), number of florets (71%), stem length (69%), stem diameter (68%), and cormel diameter (158%) and were similar to those of grown from healthy corms. However, when both Trichoderma and intercropping were applied individually to healthy corms, the results were statistically similar. In both experiments, plants treated with Trichoderma and intercropped had up to 67% less corm rot disease on average than control (plants grown without Trichoderma and intercropping). In summary, Trichoderma application to soil and intercropping with members of the Asteraceae family can be effective not only for improving growth, yield and quality of cut gladiolus, but also controlling corm rot disease of gladiolus. Moreover, intercropped species can also be a source of additional income to the growers.

Mohamed Elimem, Brahim Chermiti (Tunisia) Color Preference of Frankliniella occidentalis (Pergande) (Thysanoptera; Thripidae) and Orius sp. (Hemiptera; Anthocorridae) Populations on Two Rose Varieties(pp 94-98)

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Original Research Paper: A study of the preferential choice of Frankliniella occidentalis Pergande revealed the presence of a phototropism of this pest towards the petal color of its host plant. The color of roses may influence infestation rates by Western Flower thrips. The white-cream rose variety ‘Ociane’ attracted more individuals of F. occidentalis than the red variety ‘First-red’ in the same greenhouse; 29.54 and 39.85 thrips/flower on ‘Ociane’ while only 12.40 and 29.59 thrips/flower recorded on ‘First-red’ on May 31^st and June 14^th 2007, respectively. In fact, those differences were highly significant almost throughout the entire study period. The predatory bug Orius sp. also showed a similar preference for the ‘Ociane’ than ‘First-red’, and thus followed the distribution of its prey.

Anandhi Selvarasu, Rajamani Kandhasamy (India) Intraspecific Hybridization in Glory Lily (pp 99-102)

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Research Note: Hybridization between genetically distinct populations of a single species can serve as an important stimulus for the evolution of invasiveness. Such intraspecific hybridization was examined in Gloriosa superba, a medicinal and an emerging cut flower species. The direct and reciprocal cross of the parents in the intraspecific crosses made among germplasm of G. superba resulted in more than 90% of pod set. The parent GS 09, either as male or female parent, gave maximum pod set, fresh pod weight, number of seeds per pod, fresh seed weight per pod. The F₁ hybrids recorded the maximum mean value for all the characters over the parental mean except number of leaves per plant. The means recorded were 18.28% germination, 51.41 days for germination, 11.01 cm seedling height, 6.06 cm root length, 314.03 for vigour index (VI), 1.74 cm microtuber length, 2.12 cm microtuber girth, and 1.04 g microtuber weight.

Prabhat Kumar Singh, Raghunath Sadhukhan, Kusal Roy, Hemendra Kumar Sarkar (India) Effect of EMS on Morpho-anatomical Changes in Tuberose (Polianthes tuberosa L.)(pp 103-105)

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Research Note: Tuberose (Polianthes tuberosa L.), a perennial ornamental plant of the family Agavaceae (formerly Amaryllidaceae) and a native of Mexico, is commercially cultivated in different parts of India, including West Bengal. This flower is commercially important as a loose and cut flower, and for extraction of essential oils used in the perfumery industry. The experimental materials comprised three indigenous varieties of tuberose, ‘Calcutta Double’, ‘Prajwal’ and ‘Shringar’, which are strictly vegetatively propagated. The bulbs of all three varieties were subjected to mutagenic treatments with 0.5% and 0.25% of ethyl methane sulphonate (EMS) for inducing mutations in qualitative and quantitative characters including anatomical feature of the scape (flower-bearing stalk) and stomatal features on the leaves. Immediate effects of the mutagen were evident with respect to some qualitative characters. Induced variations leading to anatomical changes were evident in all three varieties compared to the untreated control. Variation was also exhibited in the structural organization of stomata.