取食转Bt+CpTI基因棉花的棉蚜对普通草蛉生长发育及繁殖力的影响

【目的】研究取食转Bt+CpTI基因棉花的棉蚜Aphis gossypii Glover对普通草蛉Chrysopa carnea Stephens生长发育及繁殖力的影响,为转Bt基因抗虫棉花的推广和应用提供理论支持。【方法】普通草蛉取食转Bt+CpTI棉花SKG321和对照品种石远321棉花上的棉蚜后,对普通草蛉生长发育情况进行研究,并对其幼虫、蛹的存活率和成虫的羽化率及产卵量进行分析。【结果】取食转Bt+CpTI棉花上的棉蚜对普通草蛉的各龄幼虫发育历期、蛹期和产卵前期无不利影响;处理与对照间各龄幼虫存活率、蛹的存活率和成虫羽化率之间没有显著差异;取食转Bt+CpTI棉花上棉蚜的普通草蛉单雌日产卵量与对照相比没有减少,而25 d内的单雌总产卵量与对照相比差异亦不显著。【结论】取食转Bt+CpTI棉花上的棉蚜对普通草蛉的生长发育及繁殖力无负面影响。     

转Bt基因抗虫棉对中华草蛉生物学特性的影响(英文)

为进一步揭示转Bt基因抗虫棉对中华草蛉Chrysopa sinica(Tjeder)生物学特性的影响,以泗棉3号和GK-12为材料,以棉蚜为猎物,研究了转基因抗虫棉对中华草蛉存活率、发育历期和繁殖力的影响。结果表明,转基因抗虫棉对中华草蛉幼虫期存活率、幼虫发育历期、蛹发育历期与对照差异均不显著;对成虫寿命、体重以及雌虫繁殖力也无显著差异。表明转Bt抗虫棉对中华草蛉的生物学特性影响较小。     

取食经历对美洲斑潜蝇雌成虫寿命及生殖力的影响

在实验室条件下 ,研究了取食经历对美洲斑潜蝇雌Liriomyzasativae成虫寿命和生殖力的影响。结果表明 ,幼虫期在黄瓜取食而成虫期在四季豆上取食时 ,其雌成虫寿命、总产卵量和日平均产卵量均明显小于幼虫期在长豇豆上取食而成虫期在四季豆上取食时 (P <0 0 1 ) ;当幼虫期同在长豇豆上取食时 ,成虫在黄瓜上取食时的雌成虫寿命、总产卵量和日平均产卵量均明显小于成虫在四季豆取食时 (P<0 0 5或P <0 0 1 )。这说明美洲斑潜蝇幼虫和成虫取食经历对其成虫寿命和生殖力均有显著的影响     

Bt玉米(MG95)对粘虫的抗性和拒食作用

在室内以我国自主培育的转Bt基因玉米 (简称 :Bt玉米 )为材料 ,测定了Bt玉米对不同龄期 ( 1～ 4龄 )粘虫PseudaletiaseparataWalker的影响。测定指标包括 :幼虫体重、幼虫发育历期、幼虫死亡率、化蛹率、蛹重、蛹历期、羽化率、产卵量、成虫历期、卵孵化率。结果显示 :用Bt玉米喂养的 1～ 4龄幼虫在处理 6d及 1～ 3龄幼虫在处理 9d时的体重与对照相比均差异显著 ;除 1龄幼虫外 ,2～ 4龄幼虫处理 6d及 2～ 3龄幼虫处理 9d后的死亡率差异不显著 ;用Bt玉米喂养的各龄幼虫发育历期延长 ,与对照相比差异显著 ;1、2龄幼虫的化蛹率与对照相比有显著差异 ,3、4龄则差异不显著 ;1～ 4龄幼虫的蛹重、蛹历期及羽化率均差异显著 ;成虫历期仅 1龄幼虫差异显著 ;与对照相比 ,各龄粘虫的产卵数、卵孵化率均无显著差异。结果表明 ,1龄幼虫对Bt玉米最为敏感。另外 ,拒食实验结果显示 ,在实验开始后的 8,1 6,2 4,3 2及 48h ,非选择性测定的拒食率为 63 2 0 ,64 60 ,48 99,2 3 3 5和 0 5 4% ,选择性测定的拒食率为98 3 3 ,82 43 ,71 5 4,40 1 6和 0 2 7% ,总体趋势是拒食率随时间延长而降低 ,48h后Bt玉米对粘虫幼虫无拒食活性。     

一种半固体人工饲料连代饲养日本通草蛉幼虫

以棉蚜和his gossypii Glover饲养日本通草蛉Chrysoperla nipt）oensis（Okamoto）作为对照,用一种半固体人工饲料连续饲养10代日本通草蛉,对日本通草蛉各生长阶段的生长和发育指标进行观测。结果显示,用人工饲料饲养的日本通草蛉的幼虫历期为13～15d,蛹期为8d,较对照组的7d和6d分别延长6～8d和2d;用棉蚜饲养的对照组日本通草蛉幼虫累计存活率为94％,成虫获得率为87％,而人工饲料饲养连续10代的幼虫累计存活率为81％～89％,成虫获得率69％～79％。取食人工饲料的日本通草蛉成虫,在产卵前期、产卵量、产卵期及寿命方面与对照组存在显著的差异,人工饲料组草蛉的产卵前期为5．07—5．22d,而对照组草蛉的产卵前期为4．12d,人工饲料组草蛉的产卵量为190～390粒,对照组的单雌产卵量为667．2粒,人工饲料组草蛉的寿命为36—38d,对照组成虫寿命52d。以上结果表明,这种半固体人工饲料可以满足日本通草蛉幼虫生长发育基本需要,可用于日本通草蛉的室内继代饲养,但需要进一步优化配方。     

取食转基因棉的棉大卷叶螟对中华通草蛉捕食功能与生长发育的影响研究

以GK-12、新棉33B和SGK321等转基因棉和常规棉泗棉3号为材料,研究了取食转基因棉的棉大卷叶螟对中华通草蛉生长发育和捕食功能的影响。结果表明,中华通草蛉捕食转基因棉花品种上的棉大卷叶螟后,幼虫死亡率增加2.8%-8.8%、茧重降低0.12 mg-0.50 mg/头、羽化率下降0.7%-10.1%,幼虫及茧的历期与对照相比差异达到显著或极显著水平。草蛉对棉大卷叶螟的捕食功能反应符合HollingⅡ型,其捕食量和捕食效率均高于对照,处理1头卷叶螟幼虫所用时间短于对照,瞬间攻击率大于对照。研究认为,草蛉更易捕获取食转基因棉田的棉大卷叶螟,但捕食了这些猎物后对捕食者本身的生长发育带来一定影响。     

两种野生寄主对马铃薯甲虫繁殖策略及后代生长发育的影响

为明确野生寄主植物对马铃薯甲虫种群繁殖及生长发育方面的影响,本研究比较了天仙子、刺萼龙葵2种野生寄主对马铃薯甲虫产卵量、孵化率、发育历期以及种群生命参数的影响.结果表明:取食不同寄主植物的马铃薯甲虫产卵量和孵化率均有显著差异(P＜0.05),卵期无显著差异(P＞0.05),取食天仙子的雌虫产卵量显著低于取食马铃薯和刺萼龙葵雌虫产卵量;1龄幼虫的存活率有显著性差异(P＜0.05),取食天仙子的雌虫的下一代1龄幼虫存活率显著低于其他寄主植物1龄幼虫;取食不同寄主植物的下一代马铃薯甲虫种群净增值率(R0)、内禀增长率(rm)、周限增长率(λ)都有显著差异(P＜0.05),世代平均周期(T)无显著性差异(P＞0.05);取食天仙子的马铃薯甲虫下一代种群净增值率(R0)、内禀增长率(rm)、周限增长率(λ)显著小于取食马铃薯、刺萼龙葵的下一代种群.由此可知,天仙子作为马铃薯甲虫野生过渡寄主植物,对马铃薯甲虫种群繁殖策略的作用弱于马铃薯和刺萼龙葵.在马铃薯甲虫前沿分布区,应进一步加强对刺萼龙葵野生寄主的铲除,防止马铃薯甲虫种群进一步扩散,保障马铃薯产业健康发展和粮食安全.     

四种食料植物对八字地老虎生长发育和繁殖的影响

研究了油菜、甜菜、白菜和莴苣 4种食料植物对八字地老虎Agrotisc nigrumLinnaeus幼虫发育历期、蛹期、蛹重、成虫寿命和产卵量的影响。幼虫取食油菜时发育历期最短 (1 5 4d) ,蛹最重 (2 0 3 5mg) ,产卵量最多 (3 89 5粒 ) ,甜菜次之。幼虫取食莴苣时发育历期最长 ,蛹最轻 (1 3 6 1mg) ,产卵量最少(1 44 7粒 )。 4种食料植物对寄主的蛹期和成虫的寿命没有显著的影响     

条纹小斑蛾的生物学特性

条纹小斑蛾Thyrassia penangae(Moor)是乌蔹莓(Japanese cayratia)的重要害虫,在南昌1年发生4～5代,以老熟幼虫结茧越冬。由于该虫各世代总有极少部分个体进入滞育,少数进入越夏的个体1年只发生2代或3代。羽化时间多出现在上午7～10时,羽化后当日或次日下午交配,交配时间集中在下午3～6时,交配一般可持续12个h左右。成虫羽化后需取食花蜜做补充营养才能充分产卵。产配后次日即可产卵。第1代成虫常将卵数十粒聚产于幼嫩叶片的背面,以后各代主要聚产于花蕾上,平均每雌产卵量为43粒。幼虫为4龄。第1代主要取食叶芽、幼枝及嫩叶,以后各代主要取食花蕾。在自然条件下各虫态发育历期:卵为4～7d;幼虫为10～14d;非滞育的茧期(指幼虫结茧后的预蛹至成虫羽化的日期)为8～11d,越夏茧期为32～40d,越冬茧期为205～224d。成虫寿命为3～13d。     

转Bt基因抗虫棉对棉蚜的杀虫剂敏感性及解毒酶系的影响

采用浸叶法,以亲本常规棉作对照,室内测定了吡虫啉和辛硫磷对在转Bt基因棉上取食不同世代棉蚜Aphis gossypii Glover的毒力,并用酶标仪测定了转Bt基因棉对棉蚜酯酶、羧酸酯酶和乙酰胆碱酯酶活力的影响。结果表明,棉蚜对吡虫啉和辛硫磷的敏感性并未受Bt棉的影响,2种杀虫剂对分别取食Bt棉1代和多代（60代以上）的棉蚜与取食亲本常规棉棉蚜的LC₅₀值之间均无显著差异。在抗虫棉GK12上取食1代和21代的棉蚜,其酯酶、羧酸酯酶和乙酰胆碱酯酶的活力与取食亲本常规棉的棉蚜之间差异不显著。可见,转Bt基因棉对棉蚜的杀虫剂敏感性和解毒酶活力无显著影响。     

Tri-trophic interactions between Bt cotton, the herbivore Aphis gossypii Glover (Homoptera: Aphididae), and the predator Chrysopa pallens (Rambur) (Neuroptera: Chrysopidae)

Tri-trophic impacts of transgenic Bacillus thuringiensis (Bt) cotton GK12 and NuCOTN 99B were studied using a predator, the great lacewing Chrysopa pallens (Rambur), and its prey, the cotton aphid Aphis gossypii Glover, in laboratory feeding experiments. The parental nontransgenic cotton cultivar of GK12 was used as control. The predator was fed with uniform (aphids from a single cultivar) or mixed prey (aphids from the three cotton cultivars provided on alternate days). Mortality and development of the immature stages, pupal body mass, adult sex ratio, fecundity, and egg viability of C. pallens were measured. When fed GK12-originated aphid prey, pupal body mass of C. pallens was significantly higher than that of the control, more females emerged, and these females laid significantly more eggs. Other parameters were not impacted. Females emerging from larvae maintained on NuCOTN 99B-originated prey laid fewer eggs than those maintained on GK12. Other measurements did not differ significantly between the two Bt cotton cultivars. Compared with the control, mixed feeding significantly prolonged pupal development time and increased pupal body mass and percentage of females but did not affect other parameters. These results indicate that C. pallens is sensitive to aphid prey from different cotton cultivars. Transgenic Bt cotton GK12-originated aphid prey has no adverse impact on survival, development, and fecundity of C. pallens. Between the two Bt cotton cultivars, NuCOTN 99B-originated aphid prey provided to C. pallens in the larval stage may lower female fecundity. Mixed feeding of C. pallens with the two Bt cotton-originated prey and non-Bt prey may have some adverse impacts on pupal development.     

亚致死剂量阿维菌素和氟虫睛处理小菜蛾对寄生蜂菜蛾绒茧蜂生长发育的影响

分别在小菜蛾体内的菜蛾绒茧蜂处于卵期、早期幼虫和中期幼虫时,饲喂小菜蛾2龄幼虫亚致死剂量（＝LC₁₀）的阿维菌素和氟虫睛,研究上述杀虫剂处理对寄主体内菜蛾绒茧蜂结茧率和羽化率的影响。结果表明： 在菜蛾绒茧蜂处于卵期、早期幼虫和中期幼虫时,饲喂小菜蛾LC₁₀剂量阿维菌素处理的菜叶后,菜蛾绒茧蜂的结茧率分别下降26.6%,22.8%和5.8%,饲喂小菜蛾LC₁₀剂量氟虫睛处理的菜叶后,菜蛾绒茧蜂的结茧率分别下降76.9%,42.5%和18.5%。上述阿维菌素处理对菜蛾绒茧蜂成虫羽化率影响不显著,但上述氟虫睛处理可显著抑制菜蛾绒茧蜂成虫羽化率,在菜蛾绒茧蜂处于卵期、早期幼虫和中期幼虫时,饲喂小菜蛾LC₁₀ 剂量氟虫睛处理的菜叶可导致菜蛾绒茧蜂成虫羽化率分别下降53.1%,36.1%和47.8%。结果显示,即便是对寄主小菜蛾幼虫很低的剂量（LC₁₀剂量）也会显著危害小菜蛾幼虫体内的菜蛾绒茧蜂的生长发育。此外,饲喂小菜蛾幼虫亚致死剂量杀虫剂对菜蛾绒茧蜂生长发育的影响与杀虫剂种类及蛾绒茧蜂发育阶段有关。     

The prospect of using sub-lethal imidacloprid or pirimicarb and a parasitoid wasp, Lysiphlebus fabarum, simultaneously,to control Aphis gossypii on cucumber plants

The broad-spectrum insecticides greatly influence the control of cotton aphids; however, due to frequent chemical control, Aphis gossypii (Hemiptera: Aphididae) has developed resistance against several classes of synthetic insecticides. In this study, we explored the sub-lethal effects of imidacloprid and pirimicarb, two commonly used insecticides for aphid control, on a parasitoid wasp, Lysiphlebus fabarum (Marshall) (Braconidae: Aphidiinae), when simultaneously used to control melon aphid on cucumber plants, as part of a comprehensive study for integrated pest management. Bioassays of imidacloprid and pirimicarb were performed to calculate LC₅₀ with third instars of A. gossypii. The LC₅₀ of these insecticides (110.55 and 250.89 μg/lit, respectively) were used to expose the wasp larvae, pupae, and adult parasitoids on a cucumber leaf. The percent mortality, percent adult emergence, and sex ratio were calculated during each exposure test. Moreover, the body size, egg load, and mature egg size of wasps surviving the insecticide treatments, as well as the sex ratio of the second generation was evaluated. Regardless of the host aphid mortality, none of the insecticides caused mortality of larval stage of the parasitoid. The insecticide application on pupal stage revealed that the percentage of mortality, sex ratio, body size, and egg load of surviving wasps, as well as the sex ratio of their offspring was adversely affected by imidacloprid, but not by pirimicarb. The present study suggests pirimicarb as a preferred insecticide, with less harmful effects on the fitness components of L. fabarum, for integrated pest management of cotton aphids.     

Effects of GNA transgenic sugarcane on life history parameters of Parallorhogas pyralophagus (Marsh) (Hymenoptera: Braconidae), a parasitoid of Mexican rice borer

Parallorhogas pyralophagus (Marsh) is the principal parasitoid of the Mexican rice borer [Eoreuma loftini (Dyar)], the primary pest of sugarcane in south Texas. Insect resistant transgenic sugarcane producing Galanthus nivalis agglutinin (GNA) was developed to improve control of E. loftini. The present laboratory study addressed whether over two consecutive generations transgenic sugarcane delivered via artificial diet has adverse host-mediated effects on P. pyralophagus life history parameters. The results of this study showed that a number of life history parameters were affected by transgenic sugarcane, whereas others were not affected, and that effects varied between generations. In the first generation, adult longevity was increased by approximately 2 d, and cocoon to adult and egg to adult developmental times were prolonged by approximately 1 d in parasitoids exposed to transgenic sugarcane, whereas effects were not evident on adult size; egg load; egg to cocoon developmental time; rates of gain of longevity and egg load with adult size; and egg, larval, and pupal mortality. However, in the second generation, adult longevity was reduced by approximately 3 d, adult size by approximately 5%, egg load by approximately 24%, and rate of gain of longevity with adult size by approximately 21%, whereas effects were not evident on the rate of gain of egg load with adult size. It was concluded that although GNA transgenic sugarcane, ingested via E. loftini tissues, was not acutely toxic to P. pyralophagus, the sublethal effects on life history parameters measured in this study must be considered in a broader context to determine their possible ecological significance.     

Biology and thermal requirements of Trichogramma atopovirilia Oatman & Platner (Hymenoptera: Trichogrammatidae) parasitizing eggs of Diaphania hyalinata L. (Lepidoptera: Pyralidae)

The development and parasitism of Diaphania hyalinata L. eggs by Trichogramma atopovirilia Oatman & Platner and its thermal requirements were studied at the temperatures of 18, 21, 24, 27, 30 and 33 degrees C. Thirty eggs of D. hyalinata were exposed to three females of T. atopovirilia for 5h at 25 degrees C and incubated at the different temperatures. The developmental time from egg exposure to adult, parasitism viability, number of adults per parasitized host egg and progeny sex ratio were monitored. The developmental time from egg to adult emergence of the parasitoid exhibited inverse relationship to the temperature, lasting 24.12 days at 18 degrees C and 7.36 days at 33 degrees C. Parasitism viability at 24, 27 and 30 degrees C was higher than 90%. The ratio of T. atapovirilia adult produced per egg and its sex ratio were not affected when using D. hialynata as host. The lowest threshold temperature (Tb) and estimated degree-days over Tb required by T. atopovirilia to develop on eggs of D. hyalinata was 11.99 degrees C and 130.42 masculine C, respectively. Considering the temperature regimes of two areas where cucurbitaces are cultivated in Bahia State (Rio Real and Inhambupe County) it was estimated that T. atopovirilia can achieve more than 32 generation per year. The results suggest that T. atopovirilia has potential to control D. hyalinata eggs with better chance of success under temperature regimes ranging from 24 to 27 degrees C that meets the suitable field conditions for cropping cucurbitaces.     

Morphological and biological notes on Encarsia meritoria Gahan (Hymenoptera,Aphelinidae), a parasitoid of Trialeurodes vaporariorum (Westwood) (Homoptera,Aleyrodidae) new in Europe

Encarsia meritoria Gahan, a Neartic species recorded only in the USA, was found naturally occurring in Catalonia (north‐east Spain) in 1987. The morphology of immature stages, the rate of development in the range 12°C‐34°C, the longevity and fecundity at 24°C and some observations on its searching and host feeding behaviour are presented in this paper. Mean development times from egg laying to adult emergence ranged from 75 days at 12° C to 11 days at 28°C. The Lower Developmental Threshold computed from linear regression equations was 9°C. Females laid an average of 198 eggs (range 89–330) in an average of 34 days (longevity range 19–54 days). The intrinsic rate of increase at 24° C was 0.1717, slightly greater than the r_m of T. vaporariorum and smaller than the values reported for E. tricolor and E. formosa, which is not very promising for biological control. However, this may not be a definitive conclusion, because factors other than those considered in our experiments may play an important role infield conditions.     

The consequences of short-term grazing of bioactive forages on established adult and incoming larvae populations of Teladorsagia circumcincta in lambs

The objective of this study was to investigate the consequences of short-term grazing on bioactive forages on (i) the viability and fecundity of established adult Teladorsagia circumcincta population and (ii) the establishment and development of incoming T. circumcincta infective larvae. Forty-eight, parasite naive, 3-month old, grazing lambs were artificially infected with 8000 infective larvae of T. circumcincta on day 1 of the experiment. On day 21 p.i., lambs were allocated to one of three bioactive forage grazing treatments; chicory (Cichorium intybus), sulla (Hedysarum coronarium), lotus (Lotus pedunculatus), and the control grass/clover (Lolium perenne/Trifolium repens) forage. On day 28 of the experiment a second dose of 8000 T. circumcincta infective larvae was administered to the lambs to investigate the effects of forages on the ability of infective larvae to establish within the host. All animals were slaughtered for worm recovery on day 35, while liveweight gain, feacal egg counts (FEC) and total worm egg output were monitored regularly throughout the experiment. Although FEC or total egg output were similar among the groups, adult worm burdens at slaughter were significantly affected (P<0.05) by forage treatment during the 2 week grazing period. Lambs grazing chicory had the lowest adult worm burdens and significantly lower numbers of male worms compared to those grazing on grass/clover (P<0.01), while the lambs grazing on sulla or lotus had similar adult populations to grass/clover fed animals. The results from the worm recoveries of the second dose (immature worm burdens) were affected by physiologically and/or immunologically mediated mechanisms, which reduced larval establishment in all treatments. Nevertheless, immature worm burdens at slaughter were similar between chicory, sulla and grass/clover group, while the immature worm recoveries from the lotus group were significantly higher (P<0.05) compared to those from lambs grazing grass/clover. Overall, the results of the present study support the view that chicory can be a promising candidate species in pasture management practices to control T. circumcincta burdens.     

Effects of diet,temperature and photoperiod on development and survival of the bigeyed bug, <Emphasis Type="Italic">Geocoris lubra</Emphasis>

Bigeyed bugs (Geocoris spp., Hemiptera: Geocoridae) are common predators in Australian agricultural crops yet the development and reproductive biology of Australian geocorids has not been described before. Here we present the effects of diet, temperature and photoperiod on the development and survival of Geocoris lubra Kirkaldy from egg to adult. Nymphal survival of G. lubra reared on live aphids (Aphis gossypii Glover) was very low but improved slightly on a diet of Helicoverpa armigera (Hübner) eggs. Development was faster and nymphal survival improved significantly at 27 °C compared with 25 °C. Further investigation at 27 °C showed photoperiod influenced development time, but not survival of immature G. lubra. Development time was significantly longer at 10L:14D. Fecundity of first generation G. lubra was not affected by photoperiod, although egg viability was greater at 12L:12D.     

抗生素处理对感染Wolbachia的丽蚜小蜂生殖的影响

Wolbachia是一类广泛存在于节肢动物体内, 可以对寄主的生殖力及生殖行为产生影响的共生菌。用抗生素可以有效除去寄主体内的Wolbachia。本实验通过喂食浓度分别为1, 5和10 mg盐酸四环素/mL蔗糖水, 结合PCR检测丽蚜小蜂Encarsia formosa体内Wolbachia的去除效果; 解剖观察丽蚜小蜂F0代及F1, F2和F3代怀卵量和卵巢管数量, 评价Wolbachia对丽蚜小蜂生殖的影响。结果显示： 抗生素处理去除Wolbachia后F0代蜂的卵巢管数量与未处理蜂之间无显著差异（P=0.12）, 但F1, F2和F3代蜂的卵巢管均为6条, 显著少于F0代蜂的卵巢管数量（P<0.001）。抗生素处理去除Wolbachia后的F0代蜂怀卵量与未处理蜂相比显著下降, 但显著高于经抗生素处理去除Wolbachia后的F1, F2和F3代蜂怀卵量（P<0.001）, 后代（F1, F2, F3）蜂之间怀卵量无显著差异（P=0.59）。去除Wolbachia后, 丽蚜小蜂可以产生雄性后代, 但未见交配行为, 且雌蜂可不经交配而产生雌性后代。结果说明, Wolbachia不仅直接影响丽蚜小蜂的怀卵量, 而且还可以通过影响丽蚜小蜂卵巢管的发育影响丽蚜小蜂的怀卵量; 然而, 去除Wolbachia不改变雌蜂的孤雌生殖方式。     

Generation of reactive oxygen species during cryopreservation may improve Lilium × siberia pollen viability

During biotic and abiotic stress in plants, reactive oxygen species (ROS) may play two very different roles: high ROS concentrations can exacerbate damage, whereas low concentrations can activate defense responses. The aim of this study was to investigate the relationship between ROS generation and pollen viability after cryopreservation. ROS generation was detected from ‘Siberia’ (Lilium?×?siberia) pollen using flow cytometry with 2′,7′-dichlorodihydrofluorescein diacetate as a fluorescent probe. Pollen viability was determined by 2,3,5-triphenyltetrazolium chloride staining. ROS generation was slightly increased by rapid cooling (26.13?±?4.74 vs. 15.80?±?2.30 for fresh pollen) and significantly increased by vitrification (49.74?±?1.43; P?<?0.01). Pollen viabilities after rapid cooling and vitrification were significantly increased (58.88?±?3.76% and 70.35?±?2.90%, respectively) over that of fresh pollen (46.65?±?1.61%; P?<?0.01). No significant differences in ROS generation were associated with cold acclimation at different temperatures before rapid cooling. However, sharp decreases in viability were observed with cold acclimation at 4°C and ?20°C relative to rapid cooling without acclimation (P?<?0.01). We observed nonsignificant decreases in ROS generation among vitrification treatments that omitted different steps and a significant decrease when the unloading step was omitted (P?<?0.05). Pollen viabilities were significantly reduced when the loading or dehydration steps were omitted (P?<?0.01). No significant differences were observed in ROS generation or pollen viability among the treatments when 200 U/ml catalase was added to different solutions used in the vitrification process. Comprehensive analysis of all data indicated a positive correlation between ROS generation and pollen viability (r?=?0.651, P?<?0.001). Therefore, increasing ROS generation during cryopreservation may improve the viability of ‘Siberia’ pollen.