Effects of photoperiod and temperature on diapause induction in Conogethes punctiferalis (Lepidoptera: Pyralidae)

The yellow peach moth, Conogethes punctiferalis (Guenée), a multivoltine species that overwinters as diapausing larvae, is one of the most serious insect pests on maize in China. Effect of photoperiod and temperature on larval diapause was examined under empirical laboratory conditions. Short‐day treatments caused larval diapause at 25°C, and the critical photoperiod was between 12 and 13 h (or 12 h 51 min) light per day. No sensitive instar was identified for diapause induction under alternated short‐ (L : D 11 : 13 h) and long‐day (L : D 14 : 10 h) treatments at different larval stages. However, accumulative treatment of three instars and 10 d under short‐day treatment was required for the induction of 50% larval diapause. All larvae entered diapause at 20°C, whereas less than 3% did so at 30°C, irrespective of the long‐ or short‐day treatment. Furthermore, under the short‐day treatment, more than 90% of larvae went into diapause with temperatures ≤ 25°C, but less than 17% did so at 28°C. In contrast, under the long‐day treatment, less than 19% of larvae went into diapause with temperatures ≥ 23°C. The forward shift (5°C) of critical temperature under the long‐day regime demonstrated the compensatory effect of temperature and photoperiod on diapause induction. In conclusion, C. punctiferalis had a temperature‐dependent type I photoperiodic diapause response; there was no sensitive instar for diapause determination, but the photoperiodic accumulation time countermeasures both of the short‐day cycles and the number of instars exposed, and the photoperiodic diapause response, was a temperature‐compensated phenomenon.     

Effects of different light quality on growth, chlorophyll concentration and chlorophyll biosynthesis precursors of non-heading Chinese cabbage (Brassica campestris L.)

The aim of this study was to evaluate the effects of different light quality of light emitting diode (LED) on the growth, concentration of chlorophyll and chlorophyll biosynthesis precursors of non-heading Chinese cabbage (Brassica campestris L.). Seedlings of the cultivar Te Ai Qing were cultured for 28 days under 6 treatments: red light (R), blue light (B), green light (G), yellow light (Y), red plus blue light (RB) and dysprosium lamp (CK). Lighting experiments were performed under controlled conditions (photon flux density 150 μmol m^?2 s^?1; 12 h photoperiod; 18–20 °C). The fresh and dry mass were the greatest under RB, which were significantly higher than other light treatments. The fresh mass under RB was almost twice higher compared to other light treatments. Plant height was highest under R treatment and was lowest under B. RB treatment also lowered the plant height significantly. The highest soluble sugar concentration was observed under B. The soluble protein concentration was the greatest under RB. The R treatment was adverse to pigment accumulation. The concentration of photosynthetic pigments and chlorophyll biosynthesis precursors were higher under RB. The RB treatment was beneficial to pigment accumulation.     

Effect of photoperiod and cold acclimation on survival of mice in cold

Male albino mice (Swiss-Webster) were raised at 5 °C under short (8L:16D) and long (16L:8D) light periods. All mice were housed in groups of three to five individuals in plastic mouse cages (16 × 12 × 28 cm) until 42 days of age with food and water ad libitum and cold exposed to ?40 °C between 10:00 am and 4:00 pm to determine survival time or time until loss of righting response occurred (CT min). Under short photo-periods, survival time was 49.3 ± 4.4 min and under long photoperiods it was 38.7 ± 1.9 min (P < 0.05). A second group of mice was maintained from birth at thermoneutral temperature (22 °C) under constant darkness, short day lengths (4L:20D), or constant light in the same fashion as mentioned above. When exposed to ?20 °C survival time was found to be 80.0 ± 5.0 min for the animals kept in constant darkness, 61.1 ± 2.3 min for animals raised in short photo-periods (4L:20D) (P < 0.01), and 52.4 ± 2.3 min for mice raised in constant light (P < 0.05). After 30 min mean rectal temperature was 32.1 ± 0.47 °C for constant-darkness animals, 30.5 ± 0.43 °C for short-day animals (P < 0.02), and 28.5 ± 0.74 °C for animals raised in constant light (P < 0.05). After 60 min mean rectal temperatures for constant-dark, 4L:20D, and constant-light animals were compared and body temperature was found to be 23.7 ± 1.6, 17.3 ± 1.5 (P < 0.01), and 12.8 ± 0.87 °C (P < 0.05), respectively. From these data, it is obvious that photoperiod influences cold resistance at both cold and thermoneutral acclimation temperatures although when considered individually, cold acclimation enhances cold survival to a greater degree than does reduced light exposure.     

Asymbiotic seed germination and in vitro seedling development of <Emphasis Type="Italic">Cyrtopodium punctatum</Emphasis>: a propagation protocol for an endangered Florida native orchid

Cyrtopodium punctatum Lindley is an endangered epiphytic orchid restricted in the United States to southern Florida. Due to its ornamental value, the species was extensively collected from the wild during the past 100 years. Today, only a few plants remain in protected areas. As part of a conservation plan for the species, procedures for asymbiotic seed germination were developed. Five asymbiotic orchid seed germination media (PhytoTechnology Orchid Seed Sowing Medium, Knudson C, Malmgren Modified Terrestrial Orchid Medium, Vacin &; Went Modified Orchid Medium, and ½-strengh Murashige &; Skoog) were examined for their effectiveness in promoting seed germination and protocorm development under a 16/8 h L/D photoperiod and dark (0/24 h L/D). The influence of photoperiod on growth and development was also examined. Seeds were germinated under a 16/8 h, 12/12 h, 8/16 h L/D photoperiod, at 25 ± 3°C and allowed to develop in vitro for 10 weeks. After 10 weeks, developing seedlings were transferred to Sigma Phytatrays and returned to their assigned photoperiod treatments for continued seedling development for an additional 15 weeks. Highest germination occurred in 0/24 h L/D on PhytoTechnology Orchid Seed Sowing Medium and seedlings displayed more advanced development when cultured under 16/8 h L/D photoperiod after 15 weeks in Phytatrays. Thirty-five week old seedlings potted in coconut husk growing medium exhibited 90% survival following 5 weeks acclimatization to greenhouse conditions. This asymbiotic seed germination protocol for C. punctatum will facilitate future reintroduction projects involving this endangered species.     

Larval diapause of Monochamus urussovi and photoperiodic effects on larval development in tree bolts

To determine the larval diapause and the effect of photoperiod on development in Monochamus urussovi (Coleoptera: Cerambycidae), larvae were reared on Abies sachalinensis and Picea jezoensis logs and bolts. Larvae stopped developing in the final instar at 25°C and 16L : 8D (16 h light and 8 h dark) whereas an exposure to 5°C in the dark (134 days) following acclimation at 12°C under natural daylength led to adult emergence. When larvae were reared under 8L : 16D or 16L : 8D at 25°C with an intervening period of chilling at 5°C in the dark (112 days), a photoperiod of 8L : 16D induced a shorter time required for adult emergence after being returned to 25°C, and smaller adult body size than 16L : 8D.     

Germination responses of four native terrestrial orchids from south-west Western Australia to temperature and light treatments

We report an investigation into the impact of temperature and illumination on in vitro symbiotic and asymbiotic germination of the threatened taxon Caladenia huegelii, and three other orchid spp. namely—Caladenia latifolia, Microtis media and Pterostylis sanguinea, all species from south-west Western Australia, a recognized biodiversity hotspot. High symbiotic germination on oatmeal agar (OMA + fungal symbionts specific to each species) was recorded in three species in continuous dark incubation i.e. C. huegelii seeds (98 % germination at 25 °C), and M. media and P. sanguinea (93 and 98 % respectively at 20 °C). Highest symbiotic germination for C. latifolia (100 %) was observed at 15 and 20 °C under light treatment (12/12 h light/dark). Low temperature incubation (10 °C) significantly suppressed symbiotic germination/development of seedlings across all species. Asymbiotic media treatments assessed (OMA minus fungal symbionts, Pa₅ and ½ MS), failed to stimulate any germination with C. latifolia seeds at 20 °C in either light or dark treatments after an 8 week incubation period. Seeds of M. media sown onto ½ MS medium resulted in higher germination in all developmental stages (3–5) in dark treatment than OMA and Pa₅. Seeds of P. sanguinea sown onto ½ MS medium resulted in higher overall germination in all developmental stages (3–5) in light and dark incubation compared to OMA and Pa₅. OMA supported the highest asymbiotic germination (100 %) in both light and dark incubation with M. media (only to stage 3) but did not support germination and development with other spp. tested. Caladenia huegelii seeds reached developmental Stage 3 (i.e. germinated), but only on Pa₅ medium and only at a relatively low rate in either light (2.6 %) or dark (2.1 %). Germination was higher and development of seedlings faster overall in all test species in symbiotic compared with asymbiotic media treatments. P. sanguinea seeds demonstrated the best response (among species tested) to asymbiotic germination on ½ MS with 40–53 % of germinated seeds spread over developmental stages 3–5 in light or dark incubation (at 20 °C) respectively. Illumination had no effect on fungal symbiont growth across all species, however incubation temperature treatments (10, 15, 20 and 25 °C) affected fungal growth rate. Growth of the fungal symbionts of C. huegelii, M. media and C. latifolia demonstrated significantly lower activity at 10 °C, but the cumulative radial growth rate of the P. sanguinea fungal symbiont reached 64 cm² after only 2 weeks at all temperatures tested, including 10 °C. The study highlights differences in symbiotic and aysmbiotic germination and early protocorm development in vitro between co-occurring herbaceous terrestrial Australian orchid taxa in response to variations in basal media, temperature and light.     

Effect of Temperature and Kinetin on the Germination and Endogenous Hormones of Hyoscyamus muticus Seeds

Germination of Hyoscyamus muticus L. seeds was investigated under different temperature, light and kinetin treatments. Diurnal temperature fluctuation of 25°C for 14 h and 5°C for 10 h, strongly promoted germination. Kinetin (60 mg/1) substituted for the periodic temperature changes. Both kinetin and diurnal temperature fluctuation increased the IAA level in the seeds prior to germination. A natural inhibitor (R_f 0.6–0.8) did not disappear in response to either treatment.     

Incubation temperature critical to successful stimulation of in vitro zygotic embryo growth in four Australian native Cyperaceae species

Many species of Western Australian Cyperaceae (sedges) are vital components of the indigenous flora but commonly display low seed set, poor seed quality and intractable seed dormancy. We report the effects of incubation temperature and in vitro growth media on whole seed germination compared with extracted zygotic embryo growth in Tetraria capillaris, T. octandra, Lepidosperma drummondii and L. tenue. No germination was observed from intact whole seeds of all test species regardless of the treatment evaluated. In contrast, excised zygotic embryos of all study species exhibited significant increases in growth when cultured at 15°C compared to embryos incubated at 25°C; however, optimal media for embryo growth were genera specific. Extracted embryos of T. capillaris and T. octandra exhibited maximum percentage growth (30 and 40%, respectively) at 15°C on ½ MS medium with no plant growth regulators required. In the case of L. drummondii and L. tenue 1 μM thidiazuron was a necessary addition to the ½ MS medium resulting in 40 and 77% growth of embryos (at 15°C), respectively. Incubation of extracted embryos at 25°C (regardless of medium treatment) resulted in <10% embryo growth for T. octandra and L. tenue, while the remaining two species (L. drummondii, T. capillaris) showed no embryo growth at 25°C on any medium treatment.     

Benzyladenine and low temperature promote phase transition from juvenile to vegetative adult in bulblets of Lilium?×?formolongi ‘White Aga’ cultured in vitro

Scales excised from lily bulblets were cultured on MS medium supplemented with 0.044 or 4.4 μM BA in the dark for 180 days. The culture period was divided into stage 1 (day 0–30), stage 2 (day 31–90) and stage 3 (day 91–180). The scales were cultured at 25°C in stage 1, 25°C or 8°C in stage 2, and 25°C in stage 3. When the scales were cultured on medium with 4.4 μM BA at 25°C for 180 days, bulblets with and without an elongated stem were produced. The percentage of bulblets with elongated stems greatly increased when the scales had been cultured at 8°C in stage 2. On medium with 0.044 μM BA, only bulblets without elongated stems were produced. The diameter of shoot primodia significantly enlarged in bulblets produced on medium with 4.4 μM BA at 8°C in stage 2 and no such enlargement occurred under the other conditions. Nearly square parenchyma cells were observed in the non-elongated shoot primodia in the former bulblets but not in the latter. These cells changed into longitudinally rectangular ones in the internode of elongated stems. Procambium was arranged almost parallel to the shoot axis in the stem of bulblets in the medium with 4.4 μM BA, but not in the medium with 0.044 μM BA.     

Temperature-dependent wing dimorphism in a Japanese strain of tobacco thrips, Frankliniella fusca (Thysanoptera: Thripidae)

A Japanese strain of Frankliniella fusca (Hinds) was reared under 4 temperature and two photoperiodic conditions on leaves of green bean. On the basis of the ratio of fore wing length to head width (RWL) of adult thrips we defined individuals with RWL above 3.7 as macropterous forms and those with RWL below 3.5 as brachypterous forms. Percentages of macropterous individuals among females were 0 % at 18 °C, 10.6 % at 22.5 °C, 21.8 % at 25 °C, and 77.5 % at 27.5 °C, under 15L–9D. The proportion of macropters among females developing under 15L–9D was no different from that among those developing under 10L–14D. These results indicate that photoperiod does not affect wing form, and suggest that the proportion of macropterous females increases with temperature. Although development time from hatching to adult eclosion, adult pre-oviposition period, adult head width, adult longevity, total fecundity, and fecundity schedule were compared between macropterous and brachypterous females developing at 25 °C and 15L–9D, these traits were not significantly different between the wing forms. Males of this experimental strain were all brachypterous.     

Arbuscular mycorrhizae improves low temperature stress in maize via alterations in host water status and photosynthesis

The effect of arbuscular mycorrhizal (AM) fungus, Glomus etunicatum, on growth, water status, chlorophyll concentration and photosynthesis in maize (Zea mays L.) plants was investigated in pot culture under low temperature stress. The maize plants were placed in a sand and soil mixture at 25°C for 7 weeks, and then subjected to 5°C, 15°C and 25°C for 1 week. Low temperature stress decreased AM root colonization. AM symbiosis stimulated plant growth and had higher root dry weight at all temperature treatments. Mycorrhizal plants had better water status than corresponding non-mycorrhizal plants, and significant differences were found in water conservation (WC) and water use efficiency (WUE) regardless of temperature treatments. AM colonization increased the concentrations of chlorophyll a, chlorophyll b and chlorophyll a + b. The maximal fluorescence (Fm), maximum quantum efficiency of PSII primary photochemistry (Fv/Fm) and potential photochemical efficiency (Fv/Fo) were higher, but primary fluorescence (Fo) was lower in AM plants compared with non-AM plants. AM inoculation notably increased net photosynthetic rate (Pn) and transpiration rate (E) of maize plants. Mycorrhizal plants had higher stomatal conductance (g_s) than non-mycorrhizal plants with significant difference only at 5°C. Intercellular CO₂ concentration (Ci) was lower in mycorrhizal than that in non-mycorrhizal plants, especially under low temperature stress. The results indicated that AM symbiosis protect maize plants against low temperature stress through improving the water status and photosynthetic capacity.     

The germination characteristics of <Emphasis Type="Italic">Scrophularia marilandica</Emphasis> L. (Scrophulariaceae) seeds

Investigations on seeds of Scrophularia marilandica L. were undertaken to determine their germination requirements. Seeds were collected from three naturally occurring sites and one greenhouse-grown population in London, Ontario in September and October of 1997. Some were set to germinate immediately after collection; others were stored in or on soil outside and/or under controlled laboratory conditions before testing. Germination was assessed under two light/temperature regimes (35°C 14 h light, 20°C 10 h dark and 25°C 14 h light, 10°C 10 h dark), in continuous darkness, and in the presence of two germination-promoting chemicals (GA₃ and KNO₃). Fresh seeds germinated best at 35/20°C, while stored seeds germinated best at 25/10°C. No differences in percent germination were found among three seed-maturity stages. All chemical treatments, except 0.01 M KNO_3, increased percent germination. Significant differences were found both among and within sites for most chemical treatments, but exposure to 3 × 10⁻⁴ M GA₃ caused almost every seed to germinate. When compared to the control, both the gibberellic acid and the soil-storage treatments contributed to faster germination. Exposure of seeds to naturally prevailing conditions on the soil surface followed by testing under the 25/10°C regime produced the highest percent germination. No seeds germinated in the dark. In summary, seeds of S. marilandica exhibit physiological dormancy, which can be alleviated by exposure to light, after-ripening and/or cold stratification. It is probable that the differences in germination response among sites can be attributed to differences in environmental conditions during seed production. These experiments indicate that the seeds of S. marilandica must be buried shortly after dispersal in order to form a persistent seed bank.     

Development of resistance in Cronobacter sakazakii ATCC 29544 to thermal and nonthermal processes after exposure to stressing environmental conditions

Aims: The objective was to study the response of Cronobacter sakazakii ATCC 29544 cells to heat, pulsed electric fields (PEF), ultrasound under pressure (Manosonication, MS) and ultraviolet light (UV‐C) treatments after exposure to different sublethal stresses that may be encountered in food‐processing environments. Methods and Results: Cronobacter sakazakii stationary growth‐phase cells (30°C, 24 h) were exposed to acid (pH 4·5, 1 h), alkaline (pH 9·0, 1 h), osmotic (5% NaCl, 1 h), oxidative (0·5 mmol l^?1 H₂O₂, 1 h), heat (47·5°C, 1 h) and cold (4°C, 4 h) stress conditions and subjected to the subsequent challenges: heat (60°C), PEF (25 kV cm^?1, 35°C), MS (117 μm, 200 kPa, 35°C) and UV‐C light (88·55 mW cm^?2, 25°C) treatments. The inactivation kinetics of C. sakazakii by the different technologies did not change after exposure to any of the stresses. The combinations of sublethal stress and lethal treatment that were protective were: heat shock–heat, heat shock–PEF and acid pH–PEF. Conversely, the alkaline shock sensitized the cells to heat and UV‐C treatments, the osmotic shock to heat treatments and the oxidative shock to UV‐C treatments. The maximum adaptive response was observed when heat‐shocked cells were subjected to a heat treatment, increasing the time to inactivate 99·9% of the population by 1·6 times. Conclusions: Cronobacter sakazakii resistance to thermal and nonthermal preservation technologies can increase or decrease as a consequence of previous exposure to stressing conditions. Significance and Impact of the Study: The results help in understanding the physiology of the resistance of this emerging pathogen to traditional and novel preservation technologies.     

Effect of maturation conditions on light and temperature requirements during seed germination of Citrullus colocynthis from the Arabian Desert

• Seed germination of Citrullus colocynthis, as in many other species of Cucurbitaceae, is inhibited by light, particularly at low temperatures. Germination response to light and temperature has been attributed to day length and temperature during seed maturation. This study assessed the effects of these factors on the germination response of C. colocynthis to temperature and light quality.
• Ripe fruits were collected from natural habitats during December and February and germinated at three temperatures (15/25, 20/30 and 25/35 °C) in five light treatments (dark, white light and Red:Far Red (R:FR) ratios of 0.30, 0.87 and 1.19). Additionally, unripe fruits were also collected from natural habitats and completed their maturation in growth chambers under different day lengths (6, 16 and 24 h of darkness) at 10/20 °C, and in darkness at both 10/20 °C and 25/35 °C. Mature seeds of the different treatments were germinated in the same five light treatments at 15/25 °C.
• Germination was significantly higher in the dark than that in any light treatment. Seeds matured at higher temperatures (i.e. seeds from the December collection and those matured at 25/35 °C) had significantly higher germination than those matured at lower temperatures (i.e. seeds from the February collection and those matured at 10/20 °C). Dark germination was significantly higher for the December collection than for the February collection. Seeds of the two collections germinated in the dark only at 15/25 °C. However, seeds matured in a growth chamber at 10/20 °C in darkness germinated at 15/25 °C in all light treatments, except for the R:FR ratio 0.30. Seeds of the different treatments failed to germinate in FR‐rich light.
• This study demonstrates that both temperature and day length during seed maturation play significant roles in the germination response of C. colocynthis. Additionally, the dark requirement for germination is likely beneficial for species with the larger seeds, such as C. colocynthis, which produce bigger seedlings that are able to emerge from deep soils and are competitively superior under dense vegetation and resource‐limited conditions.

     

Under low irradiation,the light regime modifies growth and metabolite production in various species of microalgae

The effects of continuous light exposure (24L:0D) and a 12 h:12 h light/dark regime (12L:12D) were compared on the growth and carotenoid, protein, sugar, lipid, and fatty acid contents in Chlorella vulgaris, Nannochloropsis sp., Isochrysis galbana, and Dunaliella salina cultured in a batchwise facility. These microalgae were grown axenically under a low photon flux density (PFD) of 27 μmol photons m^?2 s^?1. C. vulgaris, Nannochloropsis sp., and I. galbana exhibited the highest cell densities when cultured under 24L:0D, whereas D. salina grew better under the alternating light/dark regime. I. galbana accumulated high levels of proteins, sugars, and lipids and exhibited the highest carotenoid content under 24L:0D. Protein production was enhanced in C. vulgaris under 24L:0D. The highest total lipid content was recorded for D. salina, reaching 74.6 % of total proteins, sugars, and lipids in cells at the stationary phase when grown under 12L:12D. The light/dark regime at low PFD was sufficient to stimulate the accumulation of monounsaturated and polyunsaturated fatty acids in all four algae. Their levels, like those of saturated fatty acids, did not differ significantly under the two light regimes. D. salina was an important source of tetradecenoic acid 14:1(n-5). Nannochloropsis sp. produced a large amount of the essential eicosapentaenoic acid, which reached 20 % of total fatty acids under 12L:12D, while I. galbana exhibited the highest level of docosahexaenoic acid, which reached 21 % under both light regimes. This study demonstrated the feasibility of culturing microalgae under low PFD in order to produce large quantities of valuable metabolites, especially various lipids with neutraceutical value.     

Influence of temperature on the biological parameters of the anholocyclic species Cinara tujafilina (Hemiptera: Aphidoidea)

Aphids are a good model to study insect reaction to habitat change. Temperature is one of the main factors that influences insects. This paper examines the influence of temperature on developmental stages, fecundity, survival rate and demographic parameters of Cinara tujafilina (Hemiptera: Aphidoidea, Lachnidae), connected with decorative plants of the Cupressaceae family. C. tujafilina was reared in a laboratory on T. orientalis at five constant temperatures of 10, 15, 20, 25 and 28°C, 70% humidity and 14L:10D. The pre-reproduction stage varied from 7 at 25°C to 19 days at 10°C. Developmental threshold was assigned at 3.5°C. The longest reproduction stage for the aphids developing was recorded at 25°C, namely 33 days, while the shortest, at the temperature of 10°C, lasted 8 days. At 25°C this species is characterised by the shortest pre-reproduction stage, the highest fecundity, the highest survival rate and the highest demographic parameters, particularly r_m (0.17). The results suggest that the optimal temperature for the species is 25°C, and indicate that climatic change will favourably influence its development and increase its role as a pest of decorative plants.     

Differential gene expression in cucumber plants in response to brief daily cold treatments

The mechanisms of plant responses to short-term cold treatments applied daily in the period of active growth remain unknown. Cucumber (Cucumis sativus L.) plants were subjected to brief drops of temperature (2 h, 12°C) at the end of each night over a 6-day period (DROP treatment) and to prolonged (6 days) cooling at 12°C (permanent low-temperature treatment, PLT). The plants exposed to cold treatments and control plants grown at 20°C were compared in terms of cold resistance and changes in gene expression. Cold resistance of plants was determined on the basis of LT₅₀ temperature. The response of cucumber genetic machinery was assessed by means of a differential display method based on polymerase chain reaction (PCR). The changes in mRNA pool in cells of cucumber plants subjected to permanent and periodic chilling were assessed after comparing the populations of PCR fragments of cDNA. In both types of chilling protocols, the cold resistance started to increase from the 2nd day of low temperature treatment. At the end of the experiment (on the 6th day), the increment in cold resistance was three times larger for DROP compared to PLT treatment. Analysis of mRNA pool showed that the numbers of amplified fragments were nearly identical in both types of low-temperature treatment. The higher level of cold resistance under DROP conditions was assumed to depend on features of metabolism.     

Successful storage of protocorm-like bodies of hybrid Cymbidium (Orchidaceae) under low temperature conditions

Low temperatures result in lower metabolic cellular activity, thus slowing down cell division and growth. This is advantageous where a plant scientist might seek to store important germplasm without the risks associated with low temperature storage. In this study, two cold temperatures above freezing, namely 4 and 10 °C, were tested to assess for how long PLBs could be preserved without a significant loss in regeneration ability (i.e., the ability to form neo-PLBs). Control treatments were cultured at 25 °C on Teixeira Cymbidium (TC) medium at a 16-h photoperiod at a photosynthetic photon flux density (PPFD) of 45 μmol m⁻² s⁻¹. For the cold treatments, each was replicated in the dark and at low light intensity (12-h photoperiod and a PPFD of 10 μmol m⁻² s⁻¹). All cultures were sub-cultured six times onto fresh medium every 60 days, for approximately 1 year. On the 7th subculture, all neo-PLBs were prepared uniformly and replated onto standard TC medium under light conditions described above for the control. 45 days after the 7th subculture and just before subcultures 1–6, the number of neo-PLBs per half-PLB was measured. The number of neo-PLBs that formed under different treatments depended strongly on the temperature and light conditions with most neo-PLBs forming under control conditions, although that number dropped significantly as the temperature was dropped to 10 °C and then even more to 4 °C, the same trend being observed when explants were cultured and subcultured under dim light, with organogenesis being more strongly negatively influenced in darkness. For all low-temperature treatments, as well as the dimmed light and darkness treatments, the number of neo-PLBs increased significantly when recultured, on the 7th subculture, onto control TC medium under control environmental conditions, almost as high as the control values. In contrast, the control values decreased, with significantly fewer neo-PLBs by the 7th subculture relative to the control, indicating that new PLBs should be induced from shoot cultures at least once a year to maintain their vitality.     

Maternal effects of photoperiods on glycogen metabolism related to induction of diapause in Cotesia vestalis (Hymenoptera: Braconidae) from Jilin, China

Most insects must accumulate glycogen before entering diapause. Photoperiod influences diapause beyond the maternal generation in Cotesia vestalis (Haliday) (Hymenoptera: Braconidae). In this study, we measured glycogen content and the activity and messenger RNA (mRNA) expression of glycogen synthase and glycogen phosphorylase at different G₀ (25 °C under light:dark photoperiods of 8L:16D, 12L:12D, and 16L:8D) and G₁ life stages (13 °C under 8L:16D). Glycogen content in G₀ and G₁ increased with shorter light periods, except for G₀ adult males, which showed no significant difference under the three photoperiods. Compared with those under 16 h, in those under 8 h light, glycogen synthase activity was significantly higher at all tested stages, except for G₀ pupa and adult male, for which it was identical; mRNA expression was higher at G₀ larva and prepupa, lower at pupa, and identical at rest stages. Glycogen phosphorylase activity was significantly higher for G₁ egg, lower at G₁ prepupa, and identical at all other stages; mRNA expression was higher at G₀ and G₁ larval stages, and similar at rest stages. These findings suggest that glycogen, being regulated by the two enzymes, may be a consequential factor in the transmission of maternal information and diapause induction of C. vestalis.     

Environmental induction of larval diapause and life-history consequences of post-diapause development in the Large Copper butterfly,Lycaena dispar (Lepidoptera: Lycaenidae)

Many insects in temperate zones withstand the adverse conditions of winter through entering diapause and the two most important environmental stimuli that induce diapause are photoperiod and ambient temperature. The Large Copper butterfly, Lycaena dispar Haworth (Lepidoptera: Lycaenidae), is a Palearctic butterfly that hibernates as larvae. Since this butterfly is a near threatened species in some regions, there has been a growing need for a standardized protocol for mass rearing of this butterfly based on the adequate knowledge of its ecology. In the present study, we first identified that L. dispar larvae were sensitive to the photoperiodic induction of diapause during their first larval instar. We then investigated to what extent the diapause-inducing effects of photoperiod could be modified by ambient temperatures in L. dispar larvae by exposing them to the range of day-lengths (L:D 14:10, 12:12, 10:14 and 8:16) at three different temperatures (15, 20 and 25 °C). All larvae were induced to enter diapause at low ambient temperature (15 °C) regardless of photoperiod, whereas most of them (86 %) exhibited direct development when temperature was high (25 °C). The photoperiodic induction of diapause was evident when day-length was shorter than 14 h at intermediate temperature (20 °C). Pre-diapause development was prolonged at low temperatures. Finally, we found that post-diapause development of L. dispar larvae was determined by both the chilling temperature experienced by diapausing larvae and the duration of larval diapause. Adult emergence was enhanced when larvae were chilled at 8 °C and when they had been under the state of diapause for 20 days before they were treated to terminate diapause.