Cloning of heat shock protein genes from the brown planthopper,Nilaparvata lugens,and the small brown planthopper,Laodelphax striatellus,and their expression in relation to thermal stress

Three heat shock protein （HSP） genes （hsp70, hsc70, hsp90） were partially cloned from the brown planthopper Nilaparvata lugens and the small brown planthopper Laodelphax striatellus （Homoptera： Delphacidae）, which are serious pests of the rice plant. Sequence comparisons at the deduced amino acid level showed that the three HSPs of planthoppers were most homologous to corresponding HSPs of dipteran and lepidopteran species. Identities of both heat shock cognate 70 and HSP90 were higher than HSP70 in both species. Identity of the HSP70 between the two planthopper species was only 81%, a value much lower than seen among fly and moth groups. Effects of heat and cold shocks were demonstrated on expression of the three hsp genes in the two planthopper species. Heat shock （40 ℃） upregulated the hsp90 level but did not change the hsc70 level in either the nymph and adult stages of either species. On the other hand, the hsp70 level was only upregulated in L. striatellus. This heat shock response was prompt and lasted only for 1 h after treatment. In contrast, cold shock at 4℃ did not change the expression levels of any hsp in either species.     

中国对虾三倍体的诱发研究 1.温度休克

温度休克中国对虾(Penaeus orientalis)的受精卵,结果表明,用0、6和9℃的低温休克,温度越低或休克时间越长,对细胞分裂的抑制作用也越强,孵化率也越低,用30和33℃的高温休克,温度越高或休克时间越长,对细胞的伤害也越大,孵化率明显降低。低温和高温休克都能诱发三倍体,三倍体的最高诱发率,低温(9℃)为43.8％,高温(30℃)为32％,并获得三倍体蚤状幼体,对虾三倍体的诱发成功,为对虾的多倍体育种提供了可能。     

Effects of Temperature Acclimation Pretreatment on the Ultrastructure of Mesophyll Cells in Young Grape Plants （Vitis vinifera L. cv. Jingxiu） Under Cross-Temperature Stresses

Leaves from annual young grape plants （Vitis vinifera L. cv. Jingxiu） were used as experimental materials. The ultrastructural characteristics of mesophyll cells in chilling-treated plants after heat acclimation （HA） and in heat-treated plants after cold acclimation （CA） were observed and compared using transmission electron microscopy. The results showed that slight injury appeared in the ultrastructure of mesophyll cells after either HA （38℃ for 10 h） or CA （8℃ for 2.5 d）, but the tolerance to subsequent extreme temperature stress was remarkably improved by HA or CA pretreatment. The increases in membrane permeability and malondialdehyde concentration under chilling （0℃） or heat （45℃） stress were markedly inhibited by HA or CA pretreatment. The mesophyll cells of plants not pretreated with HA were markedly damaged following chilling stress. The chloroplasts appeared irregular in shape, the arrangement of the stroma lamellae was disordered, and no starch granules were present. The cristae of the mitochondria were disrupted and became empty. The nucleus became irregular in shape and the nuclear membrane was digested. In contrast, the mesophyll cells of HA-pretreated plants maintained an intact ultrastructure under chilling stress. The mesophyll cells of control plants were also severely damaged under heat stress. The chloroplast became round in shape, the stroma lamellae became swollen, and the contents of vacuoles formed clumps. In the case of mitochondria of control plants subjected to heat stress, the outer envelope was digested and the cristae were disrupted and became many small vesicles. Compared with cellular organelles in control plants, those in CA plant cells always maintained an integrated state during whole heat stress, except for the chloroplasts, which became round in shape after 10 h heat stress. From these data, we suggest that the stability of mesophyll cells under chilling stress can be increased by HA pretreatment. Similarly, CA pretreatment can protect chloroplasts, mitochondria, and the nucleus against subsequent heat stress; thus, the thermoresistance of grape seedlings was improved. The results obtained in the present study are the first, to our knowledge, to offered cytological evidence of cross-adaptation to temperature stresses in grape plants.     

Predating strategy of rodents on acorns of Quercusaliena var. acuteserrata under different predating risks and fate of acorns

The effects of rodents on forest regeneration have been highlighted in many ecological studies. In 2002 and 2003, the acorns of Quercus aliena var. acuteserrata were subjected to 12 different treatments. The daily dynamics and the amount of acorns that were finally left intact, predated in situ, or removed off were examined and documented. The ratios of acorns that were infested by bugs before and after predation by rodents were carefully documented. It was found that: (1) the ratios of acorns infested by bugs before and after predation by rodents showed significant difference (P > 0.05), suggesting that rodents would not prey on acorns during the predating process if acorns had been already infested by bugs. (2) When compared with the controls, the fate of acorns could be roughly classified into four types: acorns that were simply buried or placed on black paper showed no significant differences with the controls in their response to rodents, suggesting that rodents have no sensitivity to the little change of odor resulting from burying and may be more adapted to black background. Acorns attached with strings, dyed with black ink, cut into halves, or placed on white/green/red paper were predated in situ with much lower predation rates and lasted the same duration. Compared with the controls, the acorns were removed away by rodents with the same first day rate and the removal lasted longer. The final removal rate of the acorns by rodents was increased, whereas that of predation in situ was decreased, and none were left in the spot. This may suggest that rodents in this case increased their predation risk expectation and adopted a strategy of “less predation proportion in situ”. Acorns that were burnt were not predated in situ as well as removed in the first day, but the rates were raised to the maximum in the forth and fifth day and then declined to zero. Consequently, the final ratios of predation in situ and removal considerably decreased with many acorns, nearly 50%, being left intact. It can be hypothesized that the rodents responded to the change in odor of acorns by using the strategies of “less predation proportion in situ” and “less predating activities”, thereby resulting in larger number of acorns remained intact. Acorns that were enwrapped were barely predated in situ, removed in the first day with a normal predation rate, which declined to nearly zero in the second day and then increased till it declined again from the fourth day to the final day. The final rate of predation in situ was lower than 5% without acorns remained intact, and the rate of removal of acorns showed a considerable relative increase. This suggests that when there was a change in both odor and status of acorns, rodents also adopted the “spying” and “less predation proportion in situ” strategy, Because the predation risk is critical, the sense of sight would play an important role in rodent's living. (3) Definite amounts of acorns were predated in situ, discarded, removed away, predated ex situ, lost and hoarded separately.     

Predating strategy of rodents on acorns of Quercusaliena var. acuteserrata under different predating risks and fate of acorns

The effects of rodents on forest regeneration have been highlighted in many ecological studies. In 2002 and 2003, the acorns of Quercus aliena var. acuteserrata were subjected to 12 different treatments. The daily dynamics and the amount of acorns that were finally left intact, predated in situ, or removed off were examined and documented. The ratios of acorns that were infested by bugs before and after predation by rodents were carefully documented. It was found that: (1) the ratios of acorns infested by bugs before and after predation by rodents showed significant difference (P > 0.05), suggesting that rodents would not prey on acorns during the predating process if acorns had been already infested by bugs. (2) When compared with the controls, the fate of acorns could be roughly classified into four types: acorns that were simply buried or placed on black paper showed no significant differences with the controls in their response to rodents, suggesting that rodents have no sensitivity to the little change of odor resulting from burying and may be more adapted to black background. Acorns attached with strings, dyed with black ink, cut into halves, or placed on white/green/red paper were predated in situ with much lower predation rates and lasted the same duration. Compared with the controls, the acorns were removed away by rodents with the same first day rate and the removal lasted longer. The final removal rate of the acorns by rodents was increased, whereas that of predation in situ was decreased, and none were left in the spot. This may suggest that rodents in this case increased their predation risk expectation and adopted a strategy of “less predation proportion in situ”. Acorns that were burnt were not predated in situ as well as removed in the first day, but the rates were raised to the maximum in the forth and fifth day and then declined to zero. Consequently, the final ratios of predation in situ and removal considerably decreased with many acorns, nearly 50%, being left intact. It can be hypothesized that the rodents responded to the change in odor of acorns by using the strategies of “less predation proportion in situ” and “less predating activities”, thereby resulting in larger number of acorns remained intact. Acorns that were enwrapped were barely predated in situ, removed in the first day with a normal predation rate, which declined to nearly zero in the second day and then increased till it declined again from the fourth day to the final day. The final rate of predation in situ was lower than 5% without acorns remained intact, and the rate of removal of acorns showed a considerable relative increase. This suggests that when there was a change in both odor and status of acorns, rodents also adopted the “spying” and “less predation proportion in situ” strategy, Because the predation risk is critical, the sense of sight would play an important role in rodent's living. (3) Definite amounts of acorns were predated in situ, discarded, removed away, predated ex situ, lost and hoarded separately.     

Expression of the Tribolium castaneum （Coleoptera： Tenebrionidae） hsp83 gene and its relation to oogenesis during ovarian maturation

Heat shock proteins (HSP) can protect organisms and cells from thermal damage. In this study, we cloned the full length cDNA encoding the HSP83 protein (the homologue of HSP90) of Tribolium castaneum (red flour beetle). The isolated cDNA contains the full coding sequence, a partial 5′ untranslated region of 55 bp and the complete 3′ untranslated region. We found the hsp83 gene is located on chromosome 5 of the T. castaneum genome. The predicted HSP83 protein sequence has a high similarity (on average 86.77%) with that of other insect species. The expression of the hsp83 gene in the whole body and in the ovary could be induced with heat stress (40°C for 1 h) in newly hatched (within 3 h post emergence) and mature (10 days post emergence) beetles. Under normal conditions, the hsp83 expression in the ovary is about 3-fold higher than in the whole body at both stages. No significant difference in hsp83 expression was observed between the two ovarian developmental stages regardless if the beetles were treated with heat shock or not. The expression of the HSP83 protein in the whole body could also be induced with heat stress in newly hatched and mature beetles. However, in the ovary, HSP83 was only expressed in the follicle cells of mature beetles and not in newly hatched beetles, regardless if the beetles were treated with heat shock or not. Furthermore, the females were not able to produce mature oocytes after knock-down of the hsp83 expression by injecting dsRNA. These results suggest that the HSP83 protein is involved in protection against heat stress and could be involved in oogenesis during ovarian maturation of T. castaneum.     

Comparative demography of Liriomyza sativae Blanchard （Diptera： Agromyzidae） on cucumber at seven constant temperatures

Reproduction and population parameters of vegetable leafminer, Liriomyza sativae Blanchard were measured on cucumber （Cucumis sativus L.） at seven constant temperatures （10, 15, 20, 25, 30, 35 and 40℃）. No eggs were found at 10℃ and flies died after exposure to 40℃. The significantly highest intrinsic rate of natural increase （rm）, net reproductive rate （R0） and finite rate of increase （λ） ofL. sativae were obtained at 25℃ as 0.196, 52.452, and 1.216, respectively. The above-mentioned parameters decreased at 15℃ and 135℃ and this reduction at 35℃ was strong. Doubling time （DT） varied significantly with temperature. The shortest doubling time was obtained at 25℃. Mean generation time （T） decreased significantly with increasing temperature between 15℃ and 35℃. Percentage of immature ages in the stable age distribution was more than 95% at all temperatures. Female longevity was greater than male at all temperatures. Liriomyza sativae lived for a long time at 15℃, whereas at 35℃ had lower survival rates. The effect of temperature on reproduction, especially the intrinsic rate of increase of L. sativae would be useful for predicting its longterm population fluctuation over several generations.     

Antioxidation of Anthocyanins in Photosynthesis Under High Temperature Stress

Chlorophyll fluorescence and antioxidative capability in detached leaves of the wild type Arabidopsis thaliana L.ecotypeLandsberg erecta(Ler)and three mutants deficient in anthocyanins biosynthesis(tt3,tt4,and tt3tt4)were investigatedduring treatment with temperatures ranging 25-45℃.In comparison with the wild type,chlorophyll fluorescence parametersFv/Fm,Φ_(PSII),electron transport rate(ETR),Fv/Fo and qP in three anthocyanin-deficient mutants showed a more rapidlydecreasing rate when the temperature was over 35℃.Non-photochemical quenching(NPQ)in these mutants was almostcompletely lost at 44℃,whereas the content of heat stable protein dropped and the rate of the membrane leakage increased.Fo-temperature curves were obtained by monitoring Fo levels with gradually elevated temperatures from 22℃ to 72℃ at0.5℃/min.The inflexion temperatures of Fo were 45.8℃ in Ler,45.1℃ in tt3,44.1℃ in tt4 and 42.3℃ in tt3tt4,respectively.The temperatures of maximal Fo in three mutants were 1.9-3.8℃ lower than the wild type plants.Meanwhile,three mutantshad lower activities of superoxide dismutase(SOD)and ascorbate peroxidase(APX)and an inferior scavenging capabilityto DPPH(1.1-diphenyl-2-picrylhy.drazyl)radical under heat stress,and in particular tt3tt4 had the lowest antioxidativepotential.The results of the diaminobenzidine-H_2O_2 histochemical staining showed that H_2O_2 was accumulated in theleaf vein and mesophyll cells of mutants under treatment at 40℃,and it was significantly presented in leaf cells of tt3tt4.The sensitivity of Arabidopsis anthocyanins-deficient mutants to high temperatures has revealed that anthocyanins innormal plants might provide protection from high temperature injury,by enhancing its antioxidative capability under hightemperature stress.     

Activity of Stress-related Antioxidative Enzymes in the Invasive Plant Crofton Weed （Eupatorium adenophorum）

Crofton weed is an invasive weed in southwestern China. The activities of several antioxidative enzymes involved in plant protection against oxidative stress were assayed to determine physiological aspects of the crofton weed that might render the plant vulnerable to environmental stress. Stresses imposed on crofton weed were heat （progressively increasing temperatures： 25 ℃, 30 ℃, 35 ℃, 38℃ and 42 ℃ at 24 h intervals）, cold （progressively decreasing temperatures： 25 ℃, 20 ℃, 15℃, 10 ℃ and 5℃ at 24h intervals）, and drought （without watering up to 4days）. The three stresses induced oxidative damage as evidenced by an increase in lipid peroxidation. The effect varied with the stress imposed and the length of exposure. The activity of superoxide dismutase （SOD） increased in response to all stresses but was not significantly different from the controls （P 〈 0.05） when exposed to cold stress. Catalase （CAT） activity decreased in response to heat and drought stress but increased when exposed to cold conditions. Guaiacol peroxidase （POD） and glutathione reductase （GR） activities increased in response to cold and drought but decreased in response to heat stress. The activity of ascorbata peroxidase （APX） responded differently to all three stresses. Monodehydroascorbate reductase （MDHAR） activity decreased in response to heat and drought, and slightly increased in response to the cold stress but was not significantly different from the controls （P 〈 0.05）. The activity of dehydroascorbata reductase （DHAR） increased in response to all three stresses. Taken together, the co-ordinate increase of the oxygen-detoxifying enzymes might be more effective to protect crofton weed from the accumulation of oxygen radicals at low temperatures rather than at high temperatures.     

Effects of fluctuating thermal regimes on cold survival and life history traits of the spotted wing Drosophila(Drosophila suzukii)

Drosophila suzukii is an invasive pest causing severe damages to a large panel of cultivated crops.To facili tate its biocontrol with stratcgies such as sterile or incompatible insect techniques,D.suzukid must be mass-produced and then stored and transported under low temperature.Prolonged cold exposure induces chill injuries that can be mitigated if the cold period is interrupted with short warming intervals,referred to as fluctuating thermal regimes(FTR).In this study,we tested how to optimally use FTR to extend the shelf life of D.suzukii under cold storage.Several FTR parameters were asessed:temperature(15,20,25℃),duration(0.5,1,2,3 h),and frequency(every 12,24,36,48 h)of warming intervals,in two wild-type lines and in two developmental stages(pupac and adults).Generally,FTR improved cold storage tolerance with respect to constant low temperatures(CLT).Cold mortality was lower when recovery temperature was 20℃ or higher,when duration was 2 h per day or longer,and when warming interruptions occurred frequently(every 12 or 24 h).Applying an optimized FTR protocol to adults greatly reduced cold mortality over long-term storage(up to 130 d).Consequences of FTR on fitness-related traits were also investigated.For adults,poststorage survival was unaffected by FTR,as was the case for female fecundity and male mating capacity.On the other hand,when cold storage occurred at pupal stage,postorage survival and male mating capacity were altered under CLT,but not under FTR.After storage of pupae,female fecundity was lower under FTR compared to CLT,suggesting an energy trade-off between repair of chill damages and C22 production.This study provides detailed information on the application and optimization of an FTR-based protocol for cold storage of D.suzuki that could be useful for the biocontrol of this pest.