Effects of Auxins and Media on Callus Induction of Chinese Spring Wheat （Triticum aestivum L.）

The effects of auxins and media on callus induction from the mature and immature embryos of Chinese spring wheat （Triticum aestivum L.） varieties were investigated. It was found that genotype, medium, auxin source and concentration had the significant effects on the induction of embryogenic callus, explants germination and the increment of callus fresh weight. For immature embryos cultured on MS medium, 2 mg L^-1 of 2, 4-D was optimal, and the highest frequency of embryogenic callus （33.50%） was observed. For the mature embryos on N6 medium, 4 mg L^-1 of 2, 4-D was optimal. The frequency of embryogenic callus and increment of callus fresh weight on 2, 4, 5-T media were higher than those on 2, 4-D media, and in the presence of 2, 4, 5-T the precocious germination of explants for all genotypes were significantly suppressed. These results indicated that 2, 4, 5-T was superior to 2, 4-D and NAA in the culture of immature embryos. This is the first report about the effect of 2, 4, 5-T and NAA on wheat tissue culture, particularly in comparison with 2, 4-D in detail.     

Effects of Environmental Temperature on the Regeneration Frequency of the Immature Embryos of Wheat （Trificum aestivum L.）

The immature embryos （IEs） of wheat are the most widely used tissues for in vitro culture and genetic transformation due to its high regeneration competency. However, this explant can only be maintained in 4℃ daily cooler for a short period time for its use in plant tissue culture or transformation experiments. This study aimed to investigate the effects of environmental temperature, cryopreservation storage temperature, and heat shock culture （HSC） temperature on the regeneration frequency of wheat IEs. Results indicated that environmental temperature significantly affected the induction of embryonic calli. The optimum total accumulated temperature （TAT） during the time of anthesis and sampling for regeneration of these tissues was around 280℃ for spring wheat type cv. CB037 and approximately 300℃ for winter wheat type cv. Kenong 199. Regeneration ability obviously declined when the highest environmental temperature was over 35℃ for 1 d or a high temperature between 30 and 33℃ lasted for 5 d during anthesis and sampling. This finding was verified by culturing the freshly isolated IEs under different temperatures from 29 to 37℃ in different controlled growth incubators for 5 d; the IEs almost completely lost regeneration ability when the temperature rose to 37℃. Cryopreservation of-20℃ caused the wheat samples lost ability of producing callus or embryonic callus in a few days, and cryopreservation of-10℃ more than 10 d made the regeneration potential of the tissues dramatically declined. Comparatively, the temperature that best maintained high regeneration ability was -5℃, at which the materials can be maintained for around 1 mon. In addition, the preservation of the immature samples at -5 or -10℃ inhibited the direct germination of the IEs, avoiding the embryo axis removing process. Our results are useful for ensuring that field collection and cryopreservation of the wheat IEs are done correctly to enable tissue culture and genetic transformation.     

Mitochondrial Proteomic Analysis of Cytoplasmic Male Sterility Line and Its Maintainer in Wheat （Triticum aestivum L.）

To investigate the CMS mechanism of wheat on proteomic level and find the crucial proteins which related to fertility,mitochondria was isolated from young spike of wheat by differential centrifugation and Percoll density-gradient methods.Determined by marker enzyme assays and chlorophyll content,the mainly contaminants in the spike mitochondrial fraction were caused by peroxisomes,plastids and chloroplasts after the first discontinuous Percoll density gradient centrifugation.In order to improve the purity of spike mitochondria,a second 28% Percoll self-forming density gradient centrifugation was further carried out,the result showed that the contaminants were decreased to negligible amount,meanwhile the integrity of mitochondria （88%） was improved to 90%.The spike mitochondria proteins extracted from uninucleate stage of （S）-1376A and （A）-1376B were separated by two-dimensional electrophoresis （2-DE）,and the silver stained gels were analyzed by PDQuest 2-DE software,about 326 protein spots could be visualized on the 2-DE maps,and also revealed a similar pattern between the male sterile line and its maintainer line,except 11 spots were differentially expressed.A total of five differentially expressed proteins were analyzed by matrix-assisted laser desorption ionization time of flight mass spectrometry （MALDI-TOF-MS）,three of them were identified as manganese superoxide dismutase and T5E216 following NCBInr database by the Mascot software.These results may contribute to further understanding of the mechanisms of CMS in wheat.     

The Relationship Between Developmental Accumulation of Leaf Soluble Proteins and Vernalization Response of Wheat （Triticum aestivum L.em. Thell）

The relationship between vernalization requirement and quantitative and qualitative changes in total leaf soluble proteins were determined in one spring （cv. Kohdasht） and two winter （cvs. Sardari and Norstar） cultivars of wheat （Triticum aestivum L.） exposed to 4℃. Plants were sampled on days 2, 14, 21 and 35 of exposure to 4℃. The final leaf number （FLN） was determined throughout the vernalization periods （0, 7, 14, 24, and 35 d） at 4℃. The final leaf number decreased until days 24 and 35 in Sardari and Norstar eultivars, respectively, indicating the vernalization saturation at these times. No clear changes were detected in the final leaf number of Kohdash cultivar, verifying no vernalization requirement for this spring wheat cultivar. Comparing with control, clear cold-induced 2-fold increases in proteins quantity occurred after 48 h following the 4℃-treatment in the leaves of the both winter wheat cultivars but, such response was not detected in the spring cultivar. However, the electrophoretic protein patterns showed between-cultivar and between-temperature treatment differences. With increasing exposure time to 4℃, the winter cultivars tended to produce more HMW polypeptides than the spring cultivar. Similar proteins were induced in both Sardari and Norstar winter wheat cultivars, however, the long vernalization requirement in Norstar resulted in high level and longer duration of expression of cold-induced proteins compared to Sardari with a short vernalization requirement. These observations indicate that vernalization response regulates the expression of low temperature （LT） tolerance proteins and determines the duration of expression of LT- induced proteins.     

Allelic Variation in Loci for Adaptive Response and Its Effect on Agronomical Traits in Chinese Wheat （Triticum aestivum L.）

Heading date was an important trait that decided the adaptation of wheat to environments. It was modiifed by genes involved in vernalization response, photoperiod response and development rate. In this study, four loci Xgwm261, Xgwm219, Xbarc23 and Ppd-D1 which were previously reported related to heading time were analyzed based on three groups of wheat including landraces （L）, varieties bred before 1983 （B82） and after 1983 （A83） collected from Chinese wheat growing areas. Generally, heading date of landrace was longer than that of varieties. Signiifcant differences in the heading time existed within the groups, which implied that diversiifcation selection was much helpful for adaptation in each wheat zone. Photoperiod insensitive allele Ppd-D1a was the ifrst choice for both landrace and modern varieties, which promoted the heading date about four days earlier than that of sensitive allele Ppd-D1b. The three SSR loci had different characters in the three groups. Predominant allele combination for each zone was predicted for wheat group L and A83, which made great contribution to advantageous traits. Xgwm219 was found to be signiifcantly associated with heading date in Yellow and Huai River Winter Wheat Zone （Zone II） and spike length in Middle and lower Yangtze Valley Winter Wheat Zone （Zone III）, which implied functional diversiifcation for adaption. Variation for earliness genes provided here will be helpful for whet breeding in future climatic change.     

Photosynthesis,Growth and Antioxidant Metabolism in Mustard （Brassica juncea L.） Cultivars Differing in Cadmium Tolerance

Two mustard （Brassica juncea L. Czern and Coss.） cultivars, Pusa Jai Kisan and SS2 differing in cadmium （Cd） tolerance were treated with 0, 25 and 50 umol L-1 Cd to study the physiological basis of difference in Cd tolerance. Cultivar SS2 （Cd sensitive） accumulated greater Cd in leaves than Pusa Jai Kisan （Cd tolerant）. Further, SS2 also exhibited higher contents of thiobarbituric acid reactive substances （TBARS） and H2O2 and electrolyte leakage. However, the activities of antioxidant enzymes, catalase （CAT）, ascorbate peroxidase （APX）, dehydroascorbate reduetase （DHAR）, and glutathione reductase （GR） were higher in Pusa Jai Kisan than those in SS2. Contrarily, the activity of superoxide dismutase （SOD） was higher in SS2 than that in Pusa Jai Kisan and was the greatest at 25 umol L-1 Cd. Treatment of 25 umol L-1 Cd induced the maximum activity of enzymes. However, the activity of GR increased up to 50 umol L-1 Cd in both the cultivars. The non-enzymatic antioxidants ascorbate （AsA） and glutathione （GSH） were higher in Pusa Jai Kisan than that in SS2, whereas dehydroascorbate （DHA） and oxidized glutathione （GSSG） were higher in SS2. Photosynthesis and growth were adversely and maximally decreased by 50 umol L-1 Cd treatment in both the cultivars, but SS2 exhibited greater reductions. The protection of photosynthesis and growth and lesser reduction in Pusa Jai Kisan were associated with its capacity to restrict accumulation of Cd in leaves resulting in lower level of TBARS and H2O2 and electrolyte leakage. Moreover, Pusa Jai Kisan exhibited efficient antioxidant metabolism for removal of Cd-induced reactive oxygen species.     

Sub-Lethal Effects of Fenvalerate on the Development,Fecundity, and Juvenile Hormone Esterase Activity of Diamondback Moth,Plutella xylostella （L.）

The toxicities of fenvalerate （20% EC） to the 3rd instar larvae of diamondback moth （DBM）, Plutella xylostella （L.）, reared on three host plants viz., radish, oilseed rape, and cabbage were tested. The LC50 values of fenvalerate to the 3rd instar larvae of DBM varied with host plants, however, there wasn＇t any significant difference among them （P〉 0.05）. Similarly, DBM fed on three host plants had same pupal weight, pupal period, pupation rate, adult emergence rate, female ratio, and fecundity. The activity of juvenile hormone esterase （JHE, EC 3.1.1.1） in the 3rd instar larvae of DMB did not significantly vary with host plants, either. These suggested that DBM had similar fitness on the three host plant species. When fed on the host plants pretreated with fenvalerate at the concentrations equivalent to LC20, LC50 and LC50, the pupation rate, pupal weight, adult emergence rate, female ratio, fecundity, and JHE activity of the tested insects were declined as compared with insects in control treatments fed on the same host plant species. Furthermore, the pupal period of the tested insects was extended after fenvalerate treatment. The decrease in JHE activity after fenvalerate treatment in the tested insects could partly explain the changes in the mentioned growth parameters. Whether the role of fenvalerate in the inhibition of JHE activity could serve as a new way to control DBM needs further investigation.     

喷施海藻酸钠寡糖对小麦幼苗生长发育和抗旱性的影响

以河南省3个优质小麦品种(郑麦101、郑麦129和郑麦3596)为材料,在15%PEG6000水分胁迫条件下,研究喷施海藻酸钠寡糖对小麦幼苗生长发育和抗旱性的影响,为其作为新型植物抗旱剂在农业中的应用提供理论参考。结果表明,干旱胁迫后,3个小麦品种的生长发育均受到明显抑制,苗长、根长和生物量均显著下降,叶绿素含量也显著降低(郑麦101除外)。干旱胁迫后喷施海藻酸钠寡糖,可在一定程度上缓解干旱胁迫对小麦生长发育的抑制作用,苗长、根长和生物量均显著增加,丙二醛和脯氨酸含量显著下降,叶绿素含量(除郑麦101)和部分抗氧化酶活性有所提高。3个小麦品种相比,海藻酸钠寡糖对郑麦129和郑麦3596干旱胁迫的缓解效果好于郑麦101。     

水分与玉米秸秆还田对小麦根系生长和水分利用效率的影响

【目的】通过两年的防雨棚微区控水试验,探索秸秆还田和水分调控对小麦根系生长、产量及水分利用效率的影响,为提高秸秆还田效果及推广应用秸秆还田技术提供参考。【方法】试验设玉米秸秆粉碎翻压还田（RS）和秸秆不还田（CK）处理;3种土壤水分处理,分别为田间持水量的50%—55%（干旱处理,D）、60%—65%（轻旱处理,SD）和70%—75%（适宜水分处理,N）。测量土壤水分含量、根干重、根干重密度、根系活力、籽粒产量和水分利用效率等指标。【结果】干旱条件下小麦成熟期的次生根数显著降低,与轻旱和适宜水分处理相比,不同生育时期小麦根系活力均显著降低,0—25 cm土层中的根干重密度在不同的生育时期也基本表现为降低趋势,产量下降幅度分别为4.34%—38.30%和14.30%—36.63%,但土壤贮水消耗量分别显著增加7.92%—25.56%和31.34%—90.72%,水分利用效率分别显著增加12.69%—30.09%和11.83%—32.88%。干旱条件下,与CK处理相比,RS处理在返青期和成熟期的单株次生根数分别提高了17.17%—29.41%和5.60%—27.86%,不同生育时期0—25 cm土层中根干重密度降低,花后根系活力及25—50 cm土层中根干重密度的下降幅度增大,产量和水分利用效率分别显著降低了15.02%—19.52%和7.51%—14.56%。轻旱和适宜水分条件下,与CK处理相比,RS处理提高了不同生育时期的单株次生根数,减缓了小麦花后的根系活力及25—50 cm土层中的根干重密度下降幅度,并且增加土壤贮水消耗量,降低灌溉量及总耗水量,除2013—2014年小麦生长季适宜水分条件下不同还田方式间产量和水分利用效率差异未达显著水平外,秸秆还田处理的产量和水分利用效率分别显著提高了6.09%—9.18%和6.77%—11.13%。另外,秸秆还田方式与水分调控的交互作用显著影响小麦产量和水分利用效率。【结论】在较好的土壤水分条件下（轻旱和适宜水分）,秸秆还田对小麦根系生长具有正效应,有利于延缓根系衰老,增加土壤贮水消耗量、产量及水分利用效率,减少灌溉量;而在土壤水分条件较差时进行秸秆还田,小麦产量和水分利用效率显著降低。     

长期遮光对冬小麦群体及单叶光合特性的影响

[目的]研究长期弱光对冬小麦（Triticum aestivum L.）群体光合速率（CAP）以及上3叶特别是旗叶光合特性的影响。[方法]以耐弱光性不同的冬小麦品种扬麦158和扬麦11为材料,设不遮光（S0）、从拔节至成熟期遮去小麦冠层自然光强的22%（S1）和33%（S2）3个处理,测定小麦CAP、上3叶的光合速率（Pn）、旗叶叶绿素含量以及籽粒产量。[结果]扬麦158和扬麦11 CAP随遮光强度的增强而显著下降;长期弱光下,旗叶Pn显著下降,倒2叶Pn无明显变化,而倒3叶Pn则显著上升,从而部分地补偿了旗叶Pn的降低。该补偿效应因遮光强度和品种而异：S1处理的补偿效应大于S2处理,扬麦158的补偿效应大于扬麦11。在旗叶光合速率高值持续期内,长期遮光降低了扬麦158和扬麦11总叶绿素、叶绿素a和b的含量,以及叶绿素a/b值。[结论]弱光下,旗叶叶绿素a含量以及叶绿素a/b值下降引起其Pn显著降低,从而使得CAP明显降低,最终导致籽粒产量显著下降。该研究可为长江中下游地区的小麦生产提供重要的理论依据。     

砷对冬小麦根系生长和养分吸收的影响

[目的]研究砷对冬小麦根系生长和养分吸收的影响,揭示砷对小麦根系的毒害作用与机理。[方法]采用水培试验,设定砷浓度分别为0、5、10、15、20 mg/L,培养30 d,测定小麦根长、根体积和根系活力,分析根系中砷的累积量和养分元素含量。[结果]小麦根系能够累积较高浓度的砷,根系砷含量最高达4 506.11 mg/kg。砷显著抑制了小麦根系总吸收面积、活跃吸收面积、根长、根体积,降低了根系的生物量。溶液砷浓度为5～10 mg/L时,小麦根系中P、K、Mg和Mn、Cu、Zn的含量有增加的趋势;溶液砷浓度为15～20 mg/L时,小麦根系P、Mn、Zn的含量与对照相比均降低。小麦根系N的含量受溶液砷浓度影响不显著,砷显著降低了小麦根系Fe的含量。[结论]砷的毒害作用主要体现在抑制小麦根系生长,降低根系活力及生物量;砷对小麦根系中N、K、Ca和Mg等的含量影响较小,对Fe、Mn、Cu和Zn等微量元素含量影响较大;15～20 mg/L砷处理时,砷与P的竞争作用明显,抑制小麦根系对磷的吸收。     

The Chromosomal Control of Leaf Characteristics of Early-Stage Plants in Wheat （Triticum aestivum L.）

Rapid expansion of leaves of early-stage plants in wheat produced by chromosomal control of characteristics related to rapid expansion of the first six leaves of wheat were investigated using a set of single chromosome substitution lines under two different temperature regimes(TRs).Results from this study indicated that several chromosomes could be responsible for each of the four characteristics studied(including leaf width,leaf length,leaf elongation rate and leaf appearance index)and that different chromosomes may be responsible for the same characteristics at different TRs.Only a small number of substitutions that showed the most significant effects on leaf length were found among those that showed the most significant effects on leaf width,providing further evidence that these two characteristics are likely controlled by different sets of genes.Substitutions 4B,4D,and 5A are among those that were repeatedly detected to having significant effects on different leaf characteristics at the lower-TR and genes residing on these three chromosomes could be homologous.This possibility and its implication are discussed.     

大蒜根系水浸液处理对连作甜瓜光合及荧光特性的影响

【研究目的】为更有效的缓解甜瓜连作障碍问题,研究甜瓜与大蒜间互作效应在理论和应用上都有重要的意义。【方法】以甜瓜品种‘金蜜’为试材,盆栽于连作与非连作土壤中,用大蒜根系水浸液处理幼苗,处理后30天,测定叶绿素含量、气体交换和叶绿素荧光参数。【结果】连作使甜瓜叶片光合色素含量、净光合速率(Pn)、气孔导度(Gs)、胞间CO2浓度(Ci)、蒸腾速率(Tr)下降,使甜瓜叶片初始荧光强度(Fo)增加,PSⅡ最大光化学效率(Fv/Fm)、PSⅡ实际光化学效率(ΦPSⅡ)和光化学猝灭系数(qP)下降。大蒜根系水浸液处理连作与非连作甜瓜,均提高叶片光合色素含量、Pn、Gs、Ci、Tr、Fv/Fm、ΦPSⅡ和qP,降低Fo。【结论】甜瓜不适宜连作,大蒜根系水浸液能促进甜瓜生长,缓解甜瓜的连作障碍。     

天达-2116对冬小麦花后旗叶光合与衰老特性的调节效应

在大田条件下,研究了天达-2116对冬小麦花后旗叶光合与衰老特性的调节效应.结果表明,经天达-2116处理后,冬小麦花后旗叶叶绿素速降期的开始时间延缓,光合作用缓降期的持续时间延长,平均比CK延长了2.6 d,旗叶MDA含量升高受到抑制,叶片失水率降低,保水率增加.     

壳寡糖对PEG胁迫下小麦幼苗生长及抗氧化系统的影响

为明确壳寡糖对小麦幼苗干旱胁迫的缓解机制,采用水培试验,研究了喷施不同浓度壳寡糖溶液（10 mg/L、100 mg/L和200 mg/L）对20%PEG模拟干旱胁迫下小麦幼苗生长、叶片超氧阴离子（O·-2）和MDA含量、抗氧化酶活性以及渗透调节物质含量的影响。结果显示：喷施3种浓度壳寡糖可明显促进PEG胁迫下小麦幼苗的生长,处理48 h后幼苗株高、根长、地上部和根部干重均显著增加（200 mg/L壳寡糖对根部干重影响除外）;处理24 h和48 h后,喷施100 mg/L壳寡糖可明显降低PEG胁迫下小麦叶片的O·-2含量,而3种浓度壳寡糖均可明显降低MDA含量;相比10 mg/L和200 mg/L浓度,喷施100 mg/L壳寡糖可明显增强PEG胁迫下小麦叶片的抗氧化系统活性,SOD、POD和CAT活性及可溶性蛋白、可溶性糖和脯氨酸含量均显著提高（48 h时脯氨酸含量变化除外）。上述结果表明,100 mg/L是较适宜的喷施浓度。PEG胁迫下,喷施适宜浓度的壳寡糖能明显促进小麦地上部和根部的生长,降低叶片的活性氧含量和膜脂过氧化程度,提高抗氧化酶活性和渗透调节物质含量,增强小麦对干旱胁迫的抵抗能力。     

抗旱转基因小麦(Triticum aestivum L.) 的杂草性评价

【目的】全球转基因作物商业化迅猛发展,包括杂草化在内的一系列生态风险评估和研究尤为重要。文章中系统评价抗旱转基因小麦MGX-L大面积推广种植可能带来的演化为杂草的生态风险,为转基因抗旱小麦的环境安全评价标准制定提供科学依据。【方法】通过比较荒地和栽培地上抗旱转基因小麦、受体小麦和当地常规品种的农艺性状,评价转基因小麦的生存竞争能力;并比较荒地上生存的转基因小麦与受体的繁殖系数,评价转基因小麦群体在野外的生存能力。比较转基因小麦和受体品种的自生苗比率、种子落粒性、及深埋和浅埋地下种子萌发情况,评价转基因小麦的自然延续能力。比较转基因小麦和受体品种的花粉活力、花粉大小和花粉育性,评价转基因小麦的自然繁育能力。综合分析以上各项评价结果,评价抗旱转基因小麦演变为杂草的可能性。【结果】转基因小麦在栽培地上种植,其株高、总穗数和穗粒数与受体品种相比,没有显著差异,而转基因小麦的产量、千粒重显著高于受体品种。但和其他非转基因当地主栽小麦品种相比,或没有显著差异,或低于其他非转基因小麦品种,因此,转基因小麦与其非转基因小麦对照相似,在栽培地种植没有更强的生存竞争能力。在荒地环境下,转基因小麦生存竞争能力和受体品种没有显著差异。在荒地上种植且成熟的转基因小麦,单位面积内收获的总籽粒数少于上一年种下的种子粒数,其繁殖系数低于1。转基因小麦群体不可能在野外环境中自然维持下去。转基因小麦种子的落粒性和产生自生苗比率与受体品种相比没有显著差异。深埋20 cm以下土层的转基因小麦和受体品种的种子全部腐烂;浅埋3 cm土层的转基因小麦和受体品种的种子或者腐烂或者出苗,没有发现未萌发且保存完好能够保持活力继续发芽的种子,二者的种子腐烂或出苗的比率没有显著差异,说明转基因小麦和受体品种种子的延续能力低且无显著差异。【结论】转基因抗旱小麦MGX-L与其受体对照品种济麦22及其他非转基因当地主栽品种相似,耐旱性状的引入不会增加小麦的杂草化潜势。