Fruit, Vegetable and Cereal Science and Biotechnology

Fruit, Vegetable and Cereal Science and Biotechnology
Abbreviation: Fruit, Veg. Cereal Sci. Biotech.
Print: ISSN 1752-3419
Scope and target readership: Fruit, Vegetable and Cereal Science and Biotechnology provides a complete analysis and understanding of
any aspects of fruit, vegetable, and cereal science and biotechnology.
Fruit, Vegetable and Cereal Science and Biotechnology primarily wishes to examine the following in fruits, vegetables and cereals:
1) In vitro propagation (micropropagation, somatic embryogenesis, tissue culture, bioreactor system production), tissue culture;
2) Mycorrhizal symbioses (and effects on plant physiology, productivity, reproduction and disease resistance);
3) Cultural practices (greenhouse growth, hydroponics, aeroponics, organic farming);
4) Physiology, genetics, molecular biology, structural botany (integrated, pure and applied);
5) Pathology;
6) Phytochemistry, organic and inorganic biochemistry;
7) Storage of genetic material (cold-storage or cryopreservation) and germplasm collections (in vitro and in situ);
8) Novel techniques for analysis (genetic, biochemical, biophysical).
For publication in Fruit, Vegetable and Cereal Science and Biotechnology the research must provide a highly significant new contribution
to our understanding of fruit, vegetable and cereal crops (temperate, subtropical or tropical) and must generally be supported by a
combination of either: physiological, biochemical, genetic or molecular analyses.
Editor-in-Chief
Jaime A. Teixeira da Silva, Japan
Technical Editor
Kasumi Shima, Japan
Editorial Board and Advisory Panels
Agricultural and Agronomic Science (D1)
Emilio Cervantes, Spain
Suriyan Cha-um, Thailand
Thomas Dubois, Uganda
Hany A. El-Shemy, Egypt
Faouzi Haouala, Tunisia
Pranab Hazra, India
Xinhua He, Japan
Hashem Hussein, Egypt
Domingo Iglesias, Spain
Xinxian Li, Japan
Moahammad J. Malakouti, Iran
Reda Moghaieb, Egypt
Pio Colepicolo, Brazil
P. Ponmurugan, India
Gamal Hassan Rabie, Egypt
M. Mehdi Sharifani, Iran
Aluri Jacob Solomon Raju, India
S. Sudhakaran, India
James T. Tambong, Canada
Daniel Valero, Spain
Rajeev K. Varshney, India
Teferi Yeshitela, South Africa
Horticulture, Floriculture and Ornamental Plant
Science and Biotechnology (D11)
Chhandak Basu, USA
Emilio Cervantes, Spain
Jer-Chia Chang, Taiwan
Suriyan Cha-um, Thailand
Abdolkarim Chehregani, Iran
Samir C. Debnath, Canada
Hany A. El-Shemy, Egypt
Attila Fehér, Hungary
Manuel Fernandes-Ferreira, Portugal
C. Gopi, India
S. Dutta Gupta, India
Faouzi Haouala, Tunisia
Pranab Hazra, India
Domingo Iglesias, Spain
Sladjana Jevremovic, Serbia
Puthiyaparambil JoseKutty, New Zealand
Maurizio Lambardi, Italy
Xinxian Li, Japan
Moahammad J. Malakouti, Iran
Ezaz A. Mamun, Australia
Ákos Máthé, Hungary
Reda Moghaieb, Egypt
Benkeblia Noureddine, Japan
Suprasanna Penna, India
Edouard Pesquet, France
Pio Colepicolo, Brazil
Aluri Jacob Solomon Raju, India
Moshe Reuveni, Israel
Shyamal K. Roy, Bangladesh
Shigeru Satoh, Japan
Sanjai Saxena, India
M. Mehdi Sharifani, Iran
Dharini Sivakumar, South Africa
S. Sudhakaran, India
Judith Thomas, USA
Lining Tian, Canada
Kin-Ying To, Taiwan
Daniel Valero, Spain
Teferi Yeshitela, South Africa
Adnan Younis, Pakistan
Hao Yu, Singapore
Pathology (D13)
P. Ponmurugan, India
Gamal Hassan Rabie, Egypt
Sanjai Saxena, India
Blanca Sansegundo, Spain
Lining Tian, Canada

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Cover photos: Top left two: Redifferentiation of wheat shoots from bud clumps two weeks after transfer on regeneration
medium containing 0.1 mg/l picloram and 1 mg/l TDZ, Triticum aestivum cv. ‘Dekan’ (left); rooting of shoots on medium
with 1 mg/l IBA in barley cv. ‘Lomerit’ (Schulze, pp 64-79); Center: Tomato hydroponics in Korea (Lee et al., pp 104-109);
Bottom: screwing, a method of physically damaging apical dominance in banana (Baiyeri and Aba, pp 110-115); Top right
(2 nd from right): Bunch characters of Grand Naine banana (Musa sp. ‘AAA’) somaclonal variant GNV-04 (Phani Deepthi et
al., pp 116-120). Top right: Micropropagation of Mucuna pruriens at ILRI: rooted plantlets ready for acclimatization (Jorge
et al., pp 80-94).
Disclaimers: All comments, conclusions, opinions, and recommendations are those of the author(s), and do not necessarily
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Printed in Japan on acid-free paper.

CONTENTS
Jutta Schulze (Germany) Improvements in Cereal Tissue Culture by Thidiazuron: A Review
Maria Alexandra Jorge (Ethiopia), Marjatta Eilittä (USA), Fiona Janice Proud (Ethiopia), Barbara V. Maasdorp
(Zimbabwe), Hanna Beksissa (Ethiopia), Ashok K. Sarial (India), Jean Hanson (Ethiopia) Mucuna Species: Recent
Advances in Application of Biotechnology
Ariel R. Vicente, Gabriel O. Sozzi (Argentina) Ripening and Postharvest Storage of ‘Soft Fruits’
Guang-jae Lee, Bo-goo Kang, Tae-il Kim, Tae-jung Kim, Jin-han Kim (Korea) Tomato Hydroponics in Korea
K.P. Baiyeri, S.C. Aba (Nigeria) A Review of Protocols for Macropropagation in Musa Species
V. Phani Deepthi (India), Luke Simon (UK/India), P. Narayanaswamy (India) Identification of Elite Somaclonal Variants
from Tissue Cultured Grand Naine Banana (Musa spp. AAA) Types Using RAPDs
Julieta Andrea Silva de Almeida, Valéria Cristina Barbosa Carmazini, Luis Carlos da Silva Ramos (Brazil) Indirect Effect
of Agar Concentration on the Embryogenic Response of Coffea canephora
A. Manoj Kumar, Kalpana N. Reddy (India) Biological Control Agent in Bell Pepper Infected by Powdery Mildew
(Leveillula taurica) (Lev.) Arn.: A Biochemical Study
64
80
95
104
110
116
121
126

Fruit, Vegetable and Cereal Science and Biotechnology, VOLUME 1, NUMBER 2, 2007
Jutta Schulze (Germany) Improvements in Cereal Tissue Culture by Thidiazuron: A Review (pp 64-79)
ABSTRACT
Invited Review: This article reviews research to establish highly regenerative cell and tissue culture systems in cereals and
grasses as a prerequisite for crop improvement using biotechnological methods. The strategies were described in the historical
framework of investigations in this field and then new approaches were discussed focussing on the considerable recent
progress made using the cotton defoliant thidiazuron (TDZ) for enhancement of morphogenic competence in the Poaceae
during the last 15 years. In 1982, TDZ was first described to have cytokinin-like activity and subsequently, numerous studies
have characterized this compound as a powerful plant growth regulator for dicots with special emphasis on recalcitrant
leguminous and woody species. However, TDZ was evaluated for cereal tissue culture with a remarkable delay. The results
summarized clearly showed that TDZ can significantly improve morphogenic response from callus derived from a wide range of
explants concerning frequency of shoot formation, number of shoots per explant and the time needed for shoot induction
compared to other cytokinins. Moreover, TDZ has been used effectively for establishment of shoot meristematic cultures from
model and agronomically important cultivars using shoot apices, shoot meristematic segments as well as nodes and
pronounced differences were obtained regarding shortening of the time frame especially for barley and wheat. Besides that,
high frequency plant regeneration with long-term retention of morphogenecity in a relatively genotype-independent manner was
observed. Nevertheless, in a few cases problems associated with the use of TDZ were noted. The data summarized imply that
TDZ is also a potent growth regulator for cereals which (i) further minimizes the recalcitrant nature of the Poaceae and which (ii)
extends the application of transformation protocols to elite genotypes and to more readily available explants.
Maria Alexandra Jorge (Ethiopia), Marjatta Eilittä (USA), Fiona Janice Proud (Ethiopia), Barbara V. Maasdorp
(Zimbabwe), Hanna Beksissa (Ethiopia), Ashok K. Sarial (India), Jean Hanson (Ethiopia) Mucuna Species: Recent
Advances in Application of Biotechnology (pp 80-94)
ABSTRACT
Invited Review: Biotechnology techniques have been widely used for legumes – important crops with excellent nutritional
characteristics and soil improvement qualities. Limited work has been carried out with underutilized legume crops such as
velvet bean (Mucuna spp.), which have great potential for multiple uses. In recent decades, there has been increased interest in
the potential of Mucuna spp. as a cover crop and green manure for tropical and subtropical regions. Mucuna is also used as a
minor food crop in many countries and interest in it as livestock feed – a common use for it in the early 1900s in the USA and
elsewhere – is growing. Other minor uses exist, such as roasting the seeds as a coffee substitute. More importantly, L-dopa,
extracted from Mucuna bean seeds and plants, is used for symptomatic relief of Parkinson’s disease. Despite these numerous
qualities, some constraints have limited its adoption. Biotechnology techniques can provide a window of opportunity for new or
expanding products of Mucuna. Earlier biotechnology work with Mucuna was mostly related to its medicinal uses and focused
on the mechanism of L-dopa production. More recently, biotechnology has also been applied to identify the major virus diseases
affecting Mucuna, to develop new diagnostic methods for early virus indexing of in vitro plants and to clean virus diseases using
meristem and thermotherapy techniques, as well as to study genetic diversity through the use of molecular tools. There are still
niches to be explored such as the numerous phytochemical qualities of Mucuna that can be used to benefit human and animal
nutrition and health as well as the environment through use of these compounds in natural weed and pest control management.
Ariel R. Vicente, Gabriel O. Sozzi (Argentina) Ripening and Postharvest Storage of ‘Soft Fruits’ (pp 95-103)
ABSTRACT
Invited Review: The terms ‘soft fruit’ have been used to refer to different commodities including strawberries, blueberries and
several species of the genus Rubus. Most work in ‘soft fruit’ ripening regulation has been done on strawberry. Auxin has been
shown to be a repressor of several ripening-associated genes which led to hypothesize that reduction in auxin levels in the
receptacle could activate those genes. Despite that, some evidence suggest that that is only part of the story and that other
factors are also involved in the regulation of ‘soft fruits’ ripening process. Softening is one of the most dramatic changes
observed in ‘soft fruit’ ripening. Early reports on strawberry stated that the average molecular size of hemicelluloses greatly
declines during ripening. In contrast, later work showed that pectin size is reduced while only slight depolymerization occurs in
hemicellulosic polymers. Pectin metabolism has recently shown to be a major modification accompanying softening in species
of the genus Rubus. Contrarywise, a reduction in glucan content and downshifts in hemicellulose molecular size are the main
changes observed in blueberry. Extension of ‘soft fruit’ postharvest life has been an ongoing challenge. Strategies to reduce
‘soft fruit’ losses include selection of firmer genotypes and optimum postharvest handling procedures. A single postharvest
technique is unlikely to fully control postharvest losses but new tools such as UV radiation, heat treatments or chitosan coatings
may be added to the overall management plan (cooling conditions, modified atmospheres) to further delay softening and
prevent decay losses. Biotechnology may be useful to address some of the concerns about bramble quality attributes (e.g.
increased fruit firmness, improved flavor) and engineered resistance may be a sustainable method to control Botrytis cinerea.
Guang-jae Lee, Bo-goo Kang, Tae-il Kim, Tae-jung Kim, Jin-han Kim (Korea) Tomato Hydroponics in Korea (pp 104-109)
ABSTRACT
Invited Mini-Review: Hydroponics is ideal for improving tomato quality, increasing yields, and escaping insects and/or disease.
From a view point of these advantages, a tomato hydroponic system is highly recommended. Fertilizer application dosage and
methods affect plant growth, yield and tomato quality. Potassium accelerates lycopene accumulation, and selenium increases
fruit firmness. Chlorine and silicon increase soluble solid contents. Tomatoes contain various food functional substances such

as lycopene, β-carotene, ascorbic acid, tocopherol, etc. The kinds of salts and their concentration in nutrient solution affect fruit
texture, the amount of minerals, lycopene, and food functionality. Food functionality of tomatoes is well known to consumers,
having antioxidants that prevent cancer and heart disease. In this chapter we review the benefits of hydroponics on tomato
quality, with a focus on Korea.
K.P. Baiyeri, S.C. Aba (Nigeria) A Review of Protocols for Macropropagation in Musa Species (pp 110-115)
ABSTRACT
Invited Mini-Review: Bananas and plantains (Musa spp.) are the most important tropical fruit crops. They are staple food in
most part of the humid tropics and important source of rural income for the smallholders who produce them in compound farms.
A common limiting factor to large-scale production of Musa crops and expansion of existing plantations is the difficulty in
obtaining planting materials. This is due to poor suckering ability accentuated by the strong hormone-mediated apical
dominance exerted by the main plant. Rapid production of propagating materials could be achieved through various vegetative
multiplication techniques, including micro-propagation, but micro-propagation is not an option for the traditional small-scale
farmers in the humid tropics. Therefore, several macropropagation techniques have been developed, such as field decapitation,
excised bud, and the detached corm techniques. These techniques are relatively simple and require minimum investment to set
up, and plantlets obtained thereof, have the uniformity of tissue-culture plantlets. However, rootless explants obtained through
macropropagation have lower survival rate during the acclimatization and stabilization stages in the nursery compared to
tissue-culture plantlets. Several organic nursery substrates have been developed for optimum performance of Musa explants in
the nursery. Musa plantlets require a warm, humid, and translucent nursery environment to allow the plantlets stabilize and
escape desiccation. These conditions can be met by raising plantlets under green polyethylene chamber or under palm frond
shade as commonly practiced in tropical sub-Saharan Africa. Above all, nursery substrates must be composted for at least eight
weeks before use, and rooted explants should be preferred during nursery planting. Other valuable options discussed include
nutrient, moisture and shade management.
V. Phani Deepthi (India), Luke Simon (UK/India), P. Narayanaswamy (India) Identification of Elite Somaclonal Variants from
Tissue Cultured Grand Naine Banana (Musa spp. AAA) Types Using RAPDs (pp 116-120)
ABSTRACT
Original Research Paper: In vitro propagation of Grand Naine banana (Musa sp. ‘AAA’) is gaining importance in the banana
industry. However, the occurrence of somaclonal variants is at present limiting the use of tissue cultures plants in spite of
several advantages. Hence a brief study was carried to characterize the somaclonal variants of tissue-cultured Grand Naine
from the farmers’ fields around Bangalore, India. Eleven positive variants were characterised based on their morphological and
yield characters. To confirm the variants at the DNA level, Random Amplified Polymorphic DNA (RAPD) analysis was conducted
to identify differences in the banding patterns. Forty three primers were used for the analysis of which OPF-09 differentiated the
variants and the normal Grand Naine bananas. A band size of 320 bp was produced in all the normal samples but was absent in
the variants tested. In the present study RAPD markers proved to effectively and precisely confirm the variants identified using
molecular characters. Of the eleven superior variants analysed, variants GNV-04, GNV-08 and GNV-10 showed positive
phenotypic characters which could be used in developmental programmes of Grand Naine banana.
Julieta Andrea Silva de Almeida, Valéria Cristina Barbosa Carmazini, Luis Carlos da Silva Ramos (Brazil) Indirect Effect
of Agar Concentration on the Embryogenic Response of Coffea canephora (pp 121-125)
ABSTRACT
Original Research Paper: Coffea leaf explants can present curvature of their edges after inoculation on solid culture medium,
decreasing contact with the medium and leading to a less efficient embryogenic process. In the present study greater contact
was shown between cv. ‘Robusta’ explants and the culture medium surface when an agar concentration of 3 g L -1 was used
instead of 6 g L -1 , a concentration widely used for the majority of species inoculated in vitro. In addition, the treatment with 3 g of
agar also resulted in greater efficiency of direct somatic embryogenesis of this species, verified by an increase in the number of
sides of the explants showing the formation of embryogenic structures, and also from their size and the number of embryos
formed. Thus the reduction in agar concentration allowed for greater physical contact between the edge of the explants and the
surface of the culture medium, and also favoured the efficiency of somatic embryogenesis in leaf explants of C. canephora cv.
‘Robusta 2264 Mar’.
A. Manoj Kumar, Kalpana N. Reddy (India) Biological Control Agent in Bell Pepper Infected by Powdery Mildew (Leveillula
taurica) (Lev.) Arn.: A Biochemical Study (pp 126-130)
ABSTRACT
Original Research Paper: Biological control agents and botanicals are increasingly being used as alternative strategies to
chemicals in the control of plant diseases. The effect of some of the biological control agents and botanicals viz., Trichoderma
viride, T. harizianum and Pseudomonas fluorescens IIHR + 3, emulsified neem (Azadirachta indica) oil and NSKE (neem seed
kernel extract) as foliar spray in powdery mildew infected bell pepper, grown in a polyhouse were compared with fungicides
such as dinocap ® and Contaf ® in treated and untreated – positive control (healthy) and negative control (diseased) – plants.
Estimation of biochemical changes associated with all the above treatments was assessed. Our data show that T. viride
recorded maximum levels of 1.23, 0.92, 6.75 mg g -1 at 35 DAT and 1.0, 0.71, 8.1 mg g -1 at 50 DAT in total phenolics,
ortho-dihydroxy phenolics and total soluble sugar, respectively. The next best treatment was T. harizianum. Similarly, the
maximum amount of chlorophyll a, b, and total chlorophylls was observed in T. viride- and T. harizianum-treated plants. Overall
total phenolics and ortho-dihydroxy phenolics decreased at 50 DAT more than at 35 DAT suggesting that foliar spray of

iological control agents, botanicals and fungicides could control powdery mildew disease after one or two sprays which
resulted in low levels of pathogen load and ultimately lower levels of defense-related biochemical components. Interestingly,
during this study we found increased levels of biochemical constituents at 50 DAT in T. viride and T. harizianum even though
powdery mildew was less than 5%. These fungal biological control agents may have played a vital role in enhancing the
defense related biochemical components.

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