Two experiments were conducted independently with plants of cassava (Manihot esculenta Crantz) growing in sand with nutrient solutions with four nitrate concentrations (0.5, 3, 6 or 12 mM). In leaves, nitrate-N was undetectable at the low nitrate applications; total-N, ammonium-N, amino acid-N, reduced-N and insoluble-N all increased linearly, while soluble proteins did it curvilinearly, with increasing nitrate supply. In contrast, soluble-N did not respond to N treatments. Total-N and soluble proteins, but not nitrate-N or ammonium-N, were much higher in leaves than in roots. Plants grown under severe N deficiency accumulated ammonium-N and amino acid-N in their roots. Further, plants were exposed to either 3 or 12 mM nitrate-N, and leaf activities of key N-assimilating enzymes were evaluated. Activities of nitrate reductase, glutamine synthetase, glutamate synthase and glutamate dehydrogenase were considerably lower in low nitrate supply than in high one. Despite the low nitrate reductase activity, cassava leaves showed an ability to maintain a large proportion of N in soluble proteins.

The effect of plant growth regulators (PGR), 6-benzyladenine (BA), 2,4-dichlorophenoxyacetic acid (2,4-D), α-naphthaleneacetic acid (NAA) and sugars (sucrose, maltose, glucose, fructose) on the initiation of somatic embryogenesis of Pinus nigra Arn. was investigated. Megagametophytes containing immature zygotic embryos have been used as explants. The experiments were done in the years 2000 and 2001. Higher initiation frequencies were obtained in 2001 when the zygotic embryos showed uniformity, being in the precotyledonary stage of development. Embryogenic tissue initiation occurred on all the media tested, including PGR-free medium. Relatively high initiation frequencies were obtained on media containing either NAA (9.09 %) or 2,4-D (7.14 %) alone. Somatic embryos were present as bipolar structures and showed differences in morphological features among cell lines. Plantlet regeneration occurred in cell lines containing bipolar somatic embryos composed of compact meristematic embryo "head" and suspensor organized into bundles.

Two rice (Oryza sativa L.) cultivars, Koshihikari and Pokkali, were grown in solution culture at three concentrations of NaCl or Na₂SO₄ [0 (S0), 50 (S1), and 100 (S2) mmol dm^-3] and three N contents [0.7 (N1), 7 (N2) and 14 (N3) mmol dm^-3]. Salinity significantly decreased dry matter of both cultivars. Pokkali had better growth than Koshihikari under both saline and non-saline conditions. Applications of N enhanced development of shoot dry mass under S0 and S1 treatments up to N2. Under S2, N application had no effect on shoot dry mass of both cultivars. Root dry mass of both cultivars decreased with increasing N application at S1 and S2. Shoot and root NO₃-N content in both rice cultivars increased with increasing N concentration in the nutrient solutions. The absorption of NO₃-N was less in Koshihikari than Pokkali plants, and also was much less in Cl^- than SO₄ ^2- salinity suggesting the antagonism between Cl^- and NO₃ ^-. In addition a significant negative correlation between concentrations of NO₃-N and Cl^- in the shoots or roots was observed in both cultivars

Young plants of Lotus creticus creticus growing in a hydroponic culture were submitted to 0, 70 and 140 mM NaCl treatments for 28 d and the growth and ecophysiological characteristics of these plants have been studied. The growth of Lotus plants was not affected by salinity when applied for a short period (about 15 d); however, 140 mM NaCl induced a decrease in shoot RGR at the end of the treatment. The root growth was not decreased, even it was stimulated by 140 mM NaCl. The osmotic adjustment of Lotus plants at 70 and 140 mM NaCl maintained constant pressure potential, avoiding the visual wilting. For a similar leaf water potential, cuticular transpiration of salinized plants was lower than in control plants due to the salinity effect on the cuticle. Moreover, the presence of hairy leaves (60 and 160 trichomes per mm² in young and adult leaves, respectively) allows keeping almost 81 % of sprayed water and absorbing the 9 % of the water retained, decreased the epidermal conductance to water vapour diffusion.

Two-days-old in vitro grown protonemata of Funaria hygrometrica Hedw. were treated with a mixture PbCl₂ (4 μM Pb²⁺) and CaCl₂ (16 μM Ca²⁺) (Ca+Pb) for 48 h. The results were compared with the control: distilled water (H₂O) and the solution of PbCl₂ (4 μM Pb²⁺) (Pb). Protonemata treated with Ca+Pb were longer and contained more cells than those treated with Pb. Moreover, a lower number of cells showed apical cell deformations typical for lead toxicity: swollen tips and wall thickenings at the apex. If deformations were present they were not as extended as in Pb. In comparison with the control, however, protonemata treated with Ca+Pb were shorter, contained a lower number of cells and some apical cells in this material were altered. It can be concluded that the presence of calcium partially neutralised toxic effects of lead in Funaria hygrometrica protonemata cells.

Young maize plants, grown hydroponically, were supplied with 1/10 the optimal amount of iron (0.75 mg dm^-3). Foliar treatments with solutions, containing N⁶-benzyladenine (BA), indole-3-acetic acid (IAA) or (2-chloroethyl)-trimethylammoniumchloride (CCC) were conducted after chlorosis had been well manifested. Changes in growth, chlorophyll content, rate of photosynthesis, catalase and peroxidase activities in leaves, and the contents of Fe, Cu, Zn, Mn, and P in leaves were recorded. Growth regulators improved (CCC, IAA) or aggravated (BA) the physiological state of chlorotic plants. Their effect might be explained by changes in Fe transport towards the leaves, by increased efficiency of Fe utilization, and by effects on plant metabolism not involving Fe.

The roles of glucose-6-phosphate dehydrogenase (G6PDH) in paclitaxel production were investigated in cell suspension cultures of Taxus chinensis. In the normal cultures, the trend of G6PDH activity was similar to that of cell growth. Addition of glutamate increased G6PDH activity, while dehydroepiandrosterone (DHEA) decreased G6PDH activity. In elicitor-treated cultures, cell growth was depressed, while G6PDH activity and taxol production were enhanced compared with the control. Glutamate recovered the depression of cell growth, and resulted in further increase in G6PDH activity and taxol production. Contrarily, DHEA exacerbated the depression of cell growth, and decreased G6PDH activity and taxol production induced by fungal elicítor. The results indicated that G6PDH played a critic role of taxol production by affecting cell viability.

The effect of Cd (10, 100, and 200 μM) on tissue contents of macronutrients (N, P, K, Ca, Mg) and micronutrients (Fe, Zn, Cu, Mn) was investigated in hydroponically grown soybean (Glycine max) seedlings. Concentration changes of analysed elements observed against increasing Cd accumulation indicated that acute Cd-phytotoxic effect monitored through chlorophyll content was not a consequence of nutrient deficiency.

A Pseudomonas strain MRS16 inhibited growth of different pathogenic fungi (Aspergillus sp., Fusarium oxysporum, Pythium aphanidermatum and Rhizoctonia solani) in vitro. Larger inhibition zones were obtained on nutrient agar and King's B media compared to potato dextrose agar and pigment production media. Mutants altered in production of fluorescent pigment were derived by nitrosoguanidine mutagenesis. The pigment overproducer mutant MRS16M-1 was more inhibitory whereas nonproducer mutant MRS16M-5 was less inhibitory than parent strain on nutrient agar medium. Addition of iron (100 µM FeCl₃) in the medium decreased inhibition of fungal growth, suggesting the involvement of siderophores and other antifungal secondary metabolites. Seed bacterization of two cultivars of chickpea (Cicer arietinum cvs. H8618 and C235) differing in susceptibility to wilt caused initial root and shoot stunting at 5 d of growth followed by proliferation of secondary root growth at 10 d. Coinoculation of chickpea with Pseudomonas strain MRS16 or mutants and Rhizobium sp. Cicer strain Ca181 enhanced nodulation, nitrogen fixation and plant dry mass as compared to single inoculation with Rhizobium strain under sterile conditions.

Elevated UV-B radiation (12.2 kJ m^-2 d^-1) as against the ambient level of 10 kJ m^-2 d^-1 affected flowering, productivity and biomass partitioning of green gram [Vigna radiata (L.) Wilczek cv. KM-2]. UV-B stress delayed flowering initiation and achievement of 50 % flowering, reduced flower retention by 25 %, potential yield by 18 % and all yield attributes such as pod number (25 %), pod mass (41 %), seed number (32 %) and seed mass (45 %). Harvest index and shelling percentage were also reduced by 31 and 7 %, respectively. Application of triadimefon (20 mg dm^-3) to unstressed plants accelerated flowering and enhanced flower retention (21 %), potential yield (15 %) and yield attributes (7 to 44 %). The partitioning of biomass between plant parts also showed improvement over the control plants. In UV-B-stressed plants, triadimefon treatment compensated the inhibitions to varying extents.

Soluble sugars, proline, total chlorophyll contents and electrolyte leakage were measured in two wheat (Triticum aestivum L.) cultivars KRL 1-4 and HD 2009 at different growth stages [crown root initiation (CRI), flowering, and soft dough] under short term salinity (NaCl, CaCl₂ and Na₂SO₄). In control plants sugar contents were maximum at flowering stage. Proline and sugar concentrations increased in both cultivars under salinity with a maximum increase at CRI. Electrolyte leakage increased and chlorophyll content decreased with the plant age. A sharp increase of electrolyte leakage was noticed at salinity of 10 and 15 dS m^-1 in HD 2009 and KRL 1-4, respectively. The short-term salinity at CRI stage proved more detrimental as compared to salinity at flowering and soft dough stages in term of all biochemical changes induced. In wheat, plant resistance to salinity increased with the age of plant. The cultivar KRL 1-4 performed better under salinity as compared to HD 2009.

Glutamine synthetase (GS) showed highest expression and activity in bud (youngest topmost leaf) of Camellia sinensis, lower in older leaves, while lowest activity in stem and roots. GS expression and activity was increased by ammonium and nitrate and also by cadmium and salt stress but decreased by copper, aluminum, drought, cold and heat stress.

An efficient in vitro plant regeneration system from leaves of Ophiorrhiza japonica Blume was established for the first time. Callus formation rate was more than 90.4 % from leaf segments on Murashige and Skoog (MS) supplemented with either α-naphthaleneacetic acid (NAA) alone or in combination with 6-benzyladenine (BA). The highest shoot regeneration (78.9 %) was achieved on MS medium containing 2.0 mg dm^-3 BA and 0.2 mg dm^-3 NAA, with an average of 9.4 shoots developed per leaf segment. Shoot regeneration was also improved when the leaf explants were cultured in MS basal medium supplemented with 0.5 % (m/v) polyvinylpyrrolidone (PVP). The leaf explants from seedlings with age of about 18-27 d showed the highest shoot regeneration. The regenerated shoots were rooted on half-strength basal MS medium supplemented with 0.5 mg dm^-3 indole-3-butyric acid (IBA), which averagely produced 24.8 roots per shoot. The plantlets were transferred to soil, where 100 % survived after 1 month of acclimatization.

This paper describes the effect of NaCl on the respiration of Citrus cell suspensions namely on the induction of the alternative oxidase. The exposure of two Citrus (cvs. Carvalhal tangor and Valencia late) cell suspensions to 200 or 400 mM NaCl lead to a reduction on cell respiration rates. Under these conditions, the respiration rate decreased less in the presence of KCN indicating a stimulation of the capacity of the alternative oxidase (AOX). In addition, immunoblots showed an increase on the amount of AOX protein. Antibodies raised against the Sauromatum guttatum enzyme recognized the reduced form of the enzyme near the 35 kDa band. The protein accumulation was correlated with the significantly higher AOX capacity observed for cv. Carvalhal tangor.

Multiple shoots of Spilanthes acmella Murr. were induced from nodal buds of in vivo and in vitro seedlings on Murashige and Skoog (MS) medium containing 1.0 mg dm^-3 6-benzyladenine (BA) and 0.1 mg dm^-3 α-naphthalene-acetic acid (NAA). Adventitious shoots were successfully regenerated from the leaf explants derived from the above mentioned multiple shoots. The efficiency of shoot regeneration was tested in the MS medium containing BA, kinetin, or 2-isopentenyl adenine in combination with NAA, indole-3-acetic acid (IAA), or indole-3-butyric acid (IBA) and gibberellic acid. Maximum number of shoots per explant (20 ± 0.47) was recorded with 3.0 mg dm^-3 BA and 1.0 mg dm^-3 IAA. An anatomical study confirmed shoot regeneration via direct organogenesis. About 95 % of the in vitro shoots developed roots after transfer to half strength MS medium containing 1.0 mg dm^-3 IBA. 95 % of the plantlets were successfully acclimatized and established in soil. The transplanted plantlets showed normal flowering without any morphological variation.

An efficient, highly reproducible system for plant regeneration via somatic embryogenesis was developed for Cenchrus ciliaris genotypes IG-3108 and IG-74. Explants such as seeds, shoot tip segments and immature inflorescences were cultured on Murashige and Skoog (MS) medium supplemented with 2.0-5.0 mg dm^-3 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg dm^-3 N⁶-benzyladenine (BA) for induction of callus. Callus could be successfully induced from all the three explants of both the genotypes. But the high frequency of embryogenic callus could be induced only from immature inflorescence explants. Somatic embryos were formed from nodular, hard and compact embryogenic calli when 2,4-D concentration was gradually reduced and BA concentration increased. Histological studies of somatic embryos indicated the presence of shoot apical meristem with leaf primordia. Ultrastructural details of globular and scutellar somatic embryos further validated successful induction and progression of somatic embryogenesis. Shoots were differentiated upon germination of somatic embryos on MS medium containing 2,4-D (0.25 mg dm^-3) and BA or kinetin (1-5 mg dm^-3). Roots were induced on 1/2 MS medium containing charcoal (0.8 %), and the regenerated plants transferred to pots and established in the soil showed normal growth and fertility.

Callus cultures initiated from shoot base explants of Curcuma aromatica Salisb. were maintained on Murashige and Skoog (MS) media supplemented with 2 mg dm^-3 2,4-dichlorophenoxyacetic acid alone or with 0.5 mg dm^-3 kinetin. Plantlets were regenerated from 60 and 180-d-old callus on MS media supplemented with 3 mg dm^-3 benzyladenine and 0.5 mg dm^-3 α-naphthalene acetic acid. Approximately 8-10 plantlets were produced after 30-40 d of culture per 50 mg of callus inoculated. Out of 113 regenerants analyzed 85 plants were exclusively diploid and 28 were predominantly diploid revealing presence of polyploid nuclei. Frequency of polyploid cells were more in regenerants obtained from 180-d-old callus then from 6-d-old callus which might be attributed to the ageing of callus.

The ZEITLUPE (ZTL) protein is involved in the control of circadian period, hypocotyl elongation and flowering time in Arabidopsis thaliana. The aim of the present work was the identification of the InZTL gene and localization of its mRNA in the model short-day plant Ipomoea nil. The deduced InZTL protein of 622 amino acid residues contained a LOV domain at the N-terminal part, followed by an F-box domain and six carboxy terminal kelch repeats. Amino acid sequence of InZTL showed 84 % homology with Mesembryanthemum crystallinum ZTL (McZTL) and 83 % with Arabidopsis thaliana ZTL (AtZTL). Fluorescence in situ hybridization (FISH) to InZTL mRNA showed its high accumulation in the vascular bundles as well in the guard cells of the cotyledon. Immunolocalization of ZTL protein indicated a similar distribution pattern of ZTL protein as InZTL mRNAs.

The culture of Saussurea medusa cell were cultured in an internal loop airlift bioreactor with sifter draft tube (ILABSDT) was investigated. Under the optimal culture conditions, which were inoculation size 1.5 g(d.m.) dm^-3, aeration rate 0.3 dm³(air) dm^-3(medium) min^-1, and 14 mesh sifter holes, the maximum biomass, syringin content and syringin production reached 11.7 g(d.m.) dm^-3, 17.7 mg g^-1 and 206.6 mg dm^-3, respectively. Among cell cultures in shake flask, bubble column bioreactor and ILABSDT, ILABSDT had the highest syringin productivity and reached 12.41 mg dm^-3 d^-1.

The in vitro response of sweet cherry (Prunus cerasus × P. canescens) rootstock Gisela 5 to increasing water deficit in the culture medium was studied. Water stress induced by the incorporation of 1, 2 and 4 % polyethylene glycol (PEG-8000) into the Murashige and Skoog medium was applied for 6 weeks. PEG-induced water stress reduced shoot dry mass, length, water content and relative chlorophyll content. Water stress also induced leaf necrosis without causing loss of viability in the explants. The increase in malondialdehyde content indicated oxidative stress. The activities of antioxidant enzymes, such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), peroxidase (POX) and glutathione reductase (GR) were also significantly elevated. The concentrations of K, Ca, Fe and Mn of shoots were decreased.

Thirteen flavonoid aglycons, contained in the strongly allelopathic epicuticular exudates of Dittrichia viscosa, were investigated for their effects on lettuce seedling radicle growth. Concerning radicle length and mass, variable results were obtained, with most of the substances having no effect, some being inhibitory and some even promotive. Shoot mass was slightly reduced in four cases. Seed germination rates, root hair and lateral root formation were not affected either. Three of the compounds (namely quercetin 3,3-dimethylether, naringenin and eriodictyol) induced a strong ageotropic response in radicle growth.

The effect of various hormonal combinations on callus formation and regeneration of shoot and root from leaf derived callus of Acanthophyllum sordidum Bunge ex Boiss. has been studied. Proteins and activity of antioxidant enzymes were also evaluated during shoot and root organogenesis from callus. Calli were induced from leaf explants excised from 30-d-old seedlings grown on Murashige and Skoog medium containing 4.52 µM 2,4-dichlorophenoxyacetic acid + 4.65 µM kinetin. Maximum growth of calli and the most efficient regeneration of shoots and roots occurred with 2.69 µM 1-naphthalene acetic acid (NAA), 2.69 µM NAA + 4.54 µM thidiazuron and 2.46 µM indole-3-butyric acid. Protein content decreased in calli and increased significantly during regeneration of shoots from callus. Superoxide dismutase activity decreased in calli comparing to that of seedlings, then increased in regenerated shoots and roots. High catalase activity was detected in seedlings and regenerated shoots, whereas high peroxidase activity was observed in calli and regenerated roots.

The effects of exogenous abscisic acid (ABA) on lead tolerance in rice (Oryza sativa L.) seedlings were investigated. Pre-treatment with 0.1 g m³ ABA for 2 d restricted amount of Pb translocated from roots to shoots, decreased malondialdehyde and H₂O₂ contents in leaves, and alleviated Pb-induced decrease in plant growth and leaf chlorophyll content. Further, ABA pre-treatment adjusted leaf antioxidative enzyme activities (increased ascorbate peroxidase and catalase activities while decreased superoxide dismutase activity) and so alleviated oxidative stress.

In this work the volatiles emitted from in vitro shoot-cultures and micropropagated plants of Lavandula viridis L'Hér. were characterized and compared with those obtained from the field-grown mother-plant, using headspace solid phase micro-extraction following by capillary gas chromatography coupled to mass spectrometry (HS-SPME-GC/MS). The headspace composition consisted mainly in oxygenated monoterpenes (66.7-79.2 %), where the major constituents emitted by the mature field-grown mother-plant, in vitro shoot-cultures and micropropagated plants were 1,8-cineole (74.0, 51.9 and 57.8 %) and camphor (2.9, 15.3 and 8.7 %), respectively. The headspace of in vitro shoot-cultures and micropropagated plants showed greater amount of α-pinene, camphene, β-pinene, β-selinene and selina-3,7(11)-diene, when compared with the field-grown mother-plant.

The culture vessels with multiplying shoots of Achras zapota L. on Schenk and Hildebrandt (SH) medium containing 8.88 μM 6-benzylaminopurine (BAP) with or without sucrose were kept under varied CO₂ concentrations ranging from 0.6 to 40.0 g m^-3 using different concentrations of sodium bicarbonate (NaHCO₃), sodium carbonate (Na₂CO₃), potassium bicarbonate (KHCO₃), and potassium carbonate (K₂CO₃) in small acrylic chambers. Complete absence of carbon source caused death of shoots within 20 d. Under elevated concentrations of CO₂ (10.0 and 40.0 g m^-3) the shoots grew photoautotrophically on sucrose-free medium. The growth of cultures was better at 40.0 g (CO₂) m^-3 than on 3.0 % sucrose under ambient air of growth room. However, the best response was obtained at 10.0 g (CO₂) m^-3 and 3.0 % sucrose where maximum number of shoots, shoot length, fresh and dry mass, total number of leaves and leaf area was observed.

The leaves of axenically grown Coleus blumei were inoculated with Agrobacterium rhizogenes strain A4 and hairy root were established. PCR and Southern hybridization analysis confirmed transgenic nature of hairy root clones. Cultures of normal roots, induced by α-naphthaleneacetic acid on leaf explants, and hairy roots were evaluated for growth and rosmarinic acid content. Significantly better growth and up to 2.8 higher amount of rosmarinic acid was detected by HPLC analysis in hairy root clones. Methyl jasmonate stimulated rosmarinic acid accumulation in 6 out of 11 tested clones, while yeast extract induced RA accumulation in two and diminished it in 5 out of 11 tested hairy root clones.

It was observed that the unpollinated flowers of Cymbidium pendulum (Roxb.) Sw. and C. aloifolium (L.) Sw. stayed fresh for 20 and 18 d, respectively, but attained senescence in 8 and 7 d, respectively, after pollination. The higher content of total soluble sugars, reducing sugars and free amino acids was observed in all the floral organs of pollinated flowers than in unpollinated ones. Pollination also up-regulated the activity of hydrolytic (α-amylase, β-amylase, invertase) and proteolytic enzymes (proteases) in floral organs. Amongst floral organs, the lip and perianth possessed highest contents of metabolites. Application of auxin inhibitor (0.25 µM triiodobenzoic acid) and ethylene inhibitor (0.25 µM AgNO₃) to the pollinated flowers partially prevented the process of senescence.

Changes in growth characteristics and photochemical activities inVigna unguiculata L. Walp seedlings maintained at constant temperature of 10, 20, 30 and 40 ‡C under control and ultraviolet-B enhanced radiation (UV-B) were investigated. UV-B retarded the shoot elongation and also leaf expansion to a great extent at 30 ‡C but produced only marginal changes at 20 and 40 ‡C. Similar response was also observed with respect to changes in leaf fresh and dry masses and total chlorophyll (Chl) content under these temperatures. At 10 ‡C the total Chl content was 3-fold higher under the treatment than under control conditions. In seedlings growing at 20 and 30 ‡C the overall photosynthetic electron transport (H₂O -> methyl viologen) showed a significant enhancement during the 36-h UV-B treatment and thereafter a gradual reduction. Although a similar trend was found in photosystem 1 (PS1), the inhibition even after 60 h of UV-B treatment was not statistically significant. Photosystem 2 (PS2) activity was inhibited in seedlings treated for 60 h by UV-B at 20 and 30 ‡C. However, no inhibition was observed at 40 ‡C. No detectable photochemical activity was found in seedlings grown at 10 ‡C under either control or UV-B enhanced irradiation although the chloroplasts contained Chl.

Chenopodium rubrum, a short-day plant, and C. murale, a long-day plant, were grown in vitro in continuous darkness. Control C. rubrum plants exposed to continuous darkness for 15 d at cotyledonary phase, did not flower, while 80 % of plants flowered on the medium with 5 % glucose and 10 mg dm^-3 GA₃. Control C. murale plants exposed to continuous darkness for 10 d at the age of 4^th pair of leaves, did not flower, while GA₃ (1 - 5 mg dm^-3) stimulated flowering up to 65 %.

The effects of low growth temperature on the activities of photosystems (PS) 1 and 2 and antioxidant enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR) and catalase (CAT) in leaves of various maize inbred and hybrid genotypes (parental lines, F1 hybrids, F2 and backcross generations) were investigated. Considerable decrease of the PS 2 activity (contrary to the activity of PS 1) due to low-temperature stress was observed in the majority of genotypes/generations examined. The GR, APX and SOD activities markedly increased due to chilling, whereas the activity of CAT showed lesser changes which depended on the genotype/generation analysed. The higher susceptibility of the inbred line 2013 to low temperature was transmitted to further generations in case this line was used as the maternal parent. The intraspecific variability in photosynthetic and antioxidant parameters was caused particularly by the dominance (negative or positive), however, the level of the expression of this effect often changed after low-temperature stress and was probably the cause of the increase in the positive F1 heterosis observed in this case. Other genetic effects (e.g. the additivity, and particularly the additive or dominant maternal effects) were also found to contribute to the intraspecific variability in parameters analyzed. The dominant maternal effects possibly played an important role in maintaining positive heterosis in F2 generation.