青菜(Brassica campestris spp.chinensis var.)TPS雄性不育系的转育

在甘蓝型油菜中发现一个非常优良的不育源─—TPS雄性不育系，本研究通过远缘杂交和连续回交等手段，把TPS不育胞质从甘蓝型油菜转入青菜，选育出3个回交五代的青菜TPS不育系即青1A；青2A；青3A。不育系的不育性彻底而稳定，没有发现任何低温黄化、蜜腺退化和花器畸形等不良的细胞质负效应，群体基本稳定一致，结实性状良好。现已进入杂交组合测配阶段。

杀虫剂胁迫对小白菜B. campestris体内营养物质和黄曲条跳甲P. striolata取食的影响

采用生物测定与生理生化方法,研究不同杀虫剂胁迫下小白菜B.campestris体内的蛋白质和可溶性糖含量变化,以及对黄曲条跳甲(P.striolata)取食的影响。结果显示,杀虫剂处理对小白菜体内蛋白质含量的影响不显著,但对小白菜体内可溶性糖含量的影响显著。统计分析表明,杀虫剂处理后,黄曲条跳甲对小白菜的取食与植株体内蛋白质含量呈显著的负相关(P<0.05),与小白菜的可溶性糖含量呈极显著的正相关(P<0.01)。

Effects of Nitrogen Fertilization on the Growth and N Uptake of Late-sowing Winter Oilseed Rape (Brassica napus L.)

[Objective] The aim of this work was to analyze the N fertilization on the vegetative growth and N uptake of different winter rapeseed varieties at wintering stage. [Method] In two consecutive years （2009-2011）, two winter rapeseed varieties （B. napus L.）, an early maturity variety Zhongyou 116 （ZY116） and a middle-late application maturity variety Zhongyouza 12 （ZYZ12） were employed. Field experiments with different N levels （0, 90, 180, 270, 360 kg N/hm 2 ） were designed. At the wintering stage, the dry matter weight, the nitrogen content and concentration of plants, leaf nitrate reductase activity （NRA） and seed yields were investigated. [Result] The shoot dry matter of ZY116 increased rapidly when N rate ranged from 0 to 180 kg/hm 2 , and it raised slightly when N rate ranged from 180 to 360 kg/hm 2 . The shoot dry matter of ZYZ12 were changed in a single peak curve; the peak of shoot dry matter appeared at 270 kg N/hm 2 . The N concentration and N content in shoot and root increased rapidly when the N rate changed from 90 to 180 kg/hm 2 . Moreover, the N concentration and N content root of in ZYZ12 were much higher than that of ZY116. Present study revealed that the changed trend of leaf nitrate reductase activities （NRA） were significantly increased at the N rate of 180 kg/hm 2 in ZY116 and ZYZ12 compared with the N rate of 90 kg/hm 2 in two years. [Conclusion] Optimal nitrogen application significantly increased the dry weights and N uptake at wintering stage as well as increasing the yield of winter oilseed rape.

A Genetic Linkage Map of Brassica campestris L.ssp. pekinensis (syn. B. rapa L. ssp. pekinensis)

A molecular genetic map of Chinese cabbage was constructed with a 102 recombinant inbred (RI) population from a cross of two cultivated Chinese cabbage lines 177 and 276, using AFLP and RAPD markers. 352 markers including 265 AFLP markers and 87 RAPD markers were integrated into 17 linkage groups. It covered a total of 2 665. 7 cM with an average interval of 7. 6 cM. AFLP marker is efficient for map construction while it easily forms clusters to cause big gaps in map. A total of 13.92 % abnormal segregation markers distributed in the map. The molecular genetic map is fundamental for gene localization, comparative genomics, and QTL mapping of important agronomic traits.

Differential Expression Analysis of Genie Male Sterility A/B Lines in Chinese Cabbage-Pak-Choi (Brassica campestris ssp. chinensis Makino)

To determine differential expression of genie male sterility A/B lines in Chinese cabbage-pak-choi (Brassica campestris ssp. chinensis Makino var. communis Tsen et Lee), we used the RNA fingerprinting technique, cDNA-AFLP analysis, in different developmental stages and different tissues. While no obvious differential expressions were observed in rosette leaves, florescence leaves, and scapes, some differential expressions were found in alabstrums of A/B lines and among leaves, scapes and alabstrums. We analyzed the al-abstrums collected in different developmental stages with 10 primer combinations. We got a unique band between middle size alabstrums and large alabstrums in B line in one of the ten pair primers, and in another one pair, one band reflecting a higher gene-expression level in A line than that in B line was obtained. No unique bands were found with the other primer combinations. The bands reflecting different gene-expression level were confirmed by Northern hybridization. The results indicated that cDNA-AFLP was a suitable tool for studying differential expression of genie male sterility in plants. SDS-polyacrylamide gel electrophoresis patterns of soluble proteins further verified the difference in A/B lines.

Effect of Rare Earths on Plant under Supplementary Ultraviolet-B Radiation: Ⅰ Effect of Cerium on Growth and Photosynthesis in Rape Seedlings Exposed to Supplementary Ultraviolet-B Radiation

Effect of cerium （Ce^3＋） on growth and photosynthesis in rape seedlings exposed to two levels of ultraviolet-B radiation （UV-B, 280 - 320 nm） was studied with hydroponics under laboratory conditions. The growth of rape seedlings exposed to two levels of UV-B irradiation （0.15 and 0.35 W· m^-2/T2） was both heavily restrained. The aboveground growth indices including stem （plant） height, leaf number, leaf area, leaf fresh/dry weight and stem fresh/dry weight were obviously decreased by 13.2% - 44.1% （T1） and 21 .4% - 49.3% （T2）. Compared to CK, and except active absorption area of roots, the belowground indices main root length, root volume and fresh/dry weight by 14.1% -35.6% （T1） and 20.3% - 42.6% （T2）, respectively. For Ce ＋ UV-B treatments, the aboveground and belowground growth indices were decreased by 4.1% - 23.6%, 5.2% -23.3% （Ce＋T1） and 10.8% -28.4%, 7.0% -27.8% （Ce ＋T2）, lower than those of UV-B treatments mentioned above. These results show that Ce has protective effect on plant against injury of UV-B radiation. Furthermore, the protective effect of Ce on seedlings exposed to T1 level of UV-B radiation is superior to T2 level. Chlorophyll content, net photosynthesis rate, transpiration rate, stomatal conductance and water use efficiency in UV-B treatments decrease dramatically, whereas intercellular CO2 concentration increases. Although these indices in Ce ＋ UV-B treatments decrease compared with those of CK, the decrease in Ce ＋ UV-B treatments are lower than those in UV-B treatment. This phenomenon indicates that the ecophysiological protective effect of Ce is based on improving photosynthesis in plants. The dynamic curves of photosynthesis indices show that the course of light-repair is shortened and the injury to rape seedlings by UV-B radiation stress is alleviated by Ce.

Effect of Rare Earths on Plants under Supplementary Ultraviolet-B Radiation: Ⅱ. Effect of Cerium on Antioxidant Defense System in Rape Seedlings under Supplementary Ultraviolet-B Radiation

Effects of cerium （Ce^3＋ ） on membranous protective enzymes in rape seedlings exposed to two levels of enhanced uhraviolet-B radiation （UV-B, 280 - 320 nm） were studied by hydroponics in the laboratory. The results show that the chlorophyll content decreases and membrane permeability increases in the leaves under UV-B irradiation with an intensity of 0.15 and 0.35 W·m^-2. The activities of SOD, CAT and POD are first increased and then decreased in leaves exposed to a low level of UV-B radiation. POD activity in leaves exposed to a high level of UV-B radiation is enhanced constantly. The sensitivities of these enzymes to UV-B radiation are SOD 〉 CAT 〉 POD. The injury by UV-B radiation on the functions of protective enzymes is lightened, their ability to scavenge radicals is improved, and the membrane permeability is maintained by Ce. Furthermore, the protective effect of cerium is more obvious in plants exposed to low levels of UV-B radiation than to high levels of it. Accordingly, all results prove that the protective effect of Ce on plants under UV-B radiation is realized through the protective system of plants.

Responses of antioxidant enzyme and photosynthesis in rape seedling to the combined stresses of acid rain and ultraviolet-B radiation

Effects of the simulated acid rain（AR） and ultraviolet-B（UV-B, 280-320 nm） radiation with a single or two ways simultaneously （AR ＋ UV-B） on the antioxidant enzyme and photosynthesis of the rape seedlings were investigated by the hydroponic culture. The results of static experiment indicated that the tolerance of rape seedling to single stress（AR or UV-B） is stronger than that to dual stresses（AR ＋ UV-B）. Furthermore, the dual stresses had additive effect on catalase activity, and a synergistic effect on MDA content, net photosynthesis rate, water use efficiency as well as intercellular CO2 concentration. Meanwhile, it has an independent effect on chlorophyll content, stomatal conductance, and transpiration rate as well as membrane permeability. During 64 h restoration course, the dynamic change in the curves of physiological and biochemical indices were not identical, and none of them show a simple linear variation. According to the static and dynamic experiments, it was found that a responsive sequence of catalase activity, membrane permeability, MDA content and photosynthetic characteristics to the above-mentioned stresses was as follows： AR ＋ UV-B 〉 UV-B 〉 AR.

Brassinosteroids promote seed development and physiological maturity of oilseed rape (Brassica napus L.)

Long developmental stage and late harvest time of winter rapeseed (Brassica napus L.) have great negative effects on rice planting of rice-rapeseed farming system in China. Early maturity improvement of rapeseed is necessary. ‘Zhongshuang 11’, an elite winter rapeseed cultivar, was used in consecutive field experiments during 2010-2012. At initial flowering stage, plants were consecutively sprayed with 0.1 mg/L 2-4-Epibrassinolide(BR) for 3 d. Two hundred sampling pods from different plants were randomly collected to measure seed related indexes with a 4 d interval from 7 to 47 d after peak anthesis (DAPA).Seed color turned light brown at 31 or 35 DAPA after BR treatment, seed dry weight (DWT)was increased while seed moisture content (SMC) was decreased during seed development.DWT almost reached the maximum value when SMC was 33.20% at 31 DAPA in 2010-2011 and 35.29% at 35 DAPA in 2011-2012 growing season after BR treatment. Similarly,the maximum values of standard germination test (SGT), accelerated aging test (AAT)and cold test (CT) were observed at 31 or 35 DAPA after BR treatment respectively. The high yield and seed oil content appeared at 31 or 35 DAPA accompanied with rapid decrease in total non-structural carbohydrate (TNC) in stems and leaves. Our study indicated that BR application advanced maturity of winter rapeseed by 4 to 8 days.

UV-B辐射与酸雨复合胁迫对油菜幼苗生长的影响

采用水培实验方法研究了紫外辐射(UV-B,280～320nm)与酸雨(AR)单一和复合胁迫对油菜幼苗生长的影响。实验表明,单一胁迫与复合胁迫对油菜幼苗生长有不同程度的抑制作用。在单一紫外辐射胁迫下(T1,T2),地上部分的降幅为11.8%～44.8%、21.7%～50.4%,地下部分的降幅为2.7%～34.7%、2.8%～42.8%。单一酸雨胁迫下(AR1,AR2),降幅依次为6.2%～17.7%、3.7%～10.9%,1.6%～23.7%、0.6%～10.9%。复合胁迫下降幅明显低于单一胁迫,地上部分的降幅为24.7%～50.5%(AR1+T1)、18.5%～49.2%(AR2+T1)、32.3%～67.9%(AR1+T2)、24.4%～66.5%(AR2+T2)。地下部分的降幅依次为3.6%～46.2%、3.5%～43.1%、4.0%～52.9%、3.7%～49.7%。

稀土对UV-B辐射伤害植物的影响(Ⅰ)——Ce对UV-B辐射胁迫下油菜幼苗生长的影响

采用水培实验方法研究了紫外辐射(UV-B,280～320nm)胁迫对油菜的伤害及稀土Ce对紫外辐射伤害油菜幼苗的影响。实验表明,在紫外辐射胁迫下(T1,T2),油菜幼苗生长受到严重抑制,地下部主根长下降16.0%、20.0%,根体积下降25.0%、35.0%,根鲜重及干重分别下降35.0%、43.0%及28.0%、37.0%,根系活跃吸收面积的降幅不明显(3.0%、5.0%);地上部分的茎高,株高,叶片数,叶面积,叶鲜重、干重,茎鲜重、干重等生长指标均明显低于对照植株,降幅分别为16.0%、25.0%,18.0%、24.0%,14.0%、21.0%,45.0%、50.0%,38.0%、50.0%,34.0%、50.0%,17.0%、27.0%,20.0%、28.0%。经Ce处理的油菜幼苗,上述各项生长指标的降幅均低于未用Ce处理的植株,其降幅为5.0%、7.0%,12.0%、23.0%,24.0%、28.0%,11.0%、23.0%,2.0%、3.0%(地下部);4.0%、11.0%,8.0%、12.0%,10.0%、16.0%,24.0%、29.0%,14.0%、29.0%,15.0%、28.0%,4.0%、12.0%,5.0%、16.0%(地上部)。表明Ce对紫外辐射胁迫伤害油菜幼苗有一定缓解效应。

稀土对UV-B辐射伤害植物的影响——Ⅰ.Ce(Ⅲ)对UV-B胁迫下油菜幼苗生长及光合作用影响

用水培法研究了Ce(Ⅲ)对紫外辐射(UV-B，280，320nm)胁迫下油菜幼苗生长与光合作用的影响。实验表明，在紫外辐射(T1/0．15W·m^-2和T2／0．35W·m^-2)胁迫下，油菜幼苗生长受到严重抑制。地上部茎(株)高、叶片数、叶面积、叶鲜(干)重、茎鲜(干)重等生长指标均低于对照植株13．2％～44．1％(T1)，21．4％～49．3％(T2)；地下部除根系活跃吸收面积降幅不明显外，主根长、根体积、根鲜(干)重均低于对照14．1％～35．6％(T1)，20．3％～42．6％(T2)。Ce处理油菜幼苗，上述各项指标降幅均低于未用Ce处理植株(地上部，Ce+T1降低4．1％～23．6％，Ce+T2，降低10．8％～28．4％；地下部，Ce+T1降低5．2％～23．3％，ce+T2降低7．0％～27．8％)。表明Ce对紫外辐射伤害油菜幼苗有防护效应，且对低剂量UV-B伤害的防护效果更优。光合参数测定显示，UV-B胁迫下植株Chl含量，Pn，En，Gs及WUE降低，Ci增高。Ce处理植物的5项参数降幅减小，表明对光合作用的改善是Ce防护效应的生态生理学基础。光合参数动态测定显示，Ce介导使油菜光致修复历程缩短(逼向CK的拐点前置)，或各时段变幅陡减。

转抗菌肽B基因和bar基因籼稻植株的再生

运用基因枪法将含有 bar基因和 cecropin B基因的质粒 p CB1导入籼稻品种特青的幼胚细胞 ,筛选后得到 3株转基因植株。PCR检测和 Southern杂交结果表明 ,外源基因已整合到水稻基因组中。转化当代植株表现出对 Basta很强的抗性 。

白菜雄性不育相关基因BcMF4基因功能的RNAi验证

BcMF4(Brassica campestris Male Fertility 4)是前一阶段从普通白菜(Brassica campestris ssp．chinensis var．communis,syn．B．rapa ssp．chinensis var．communis)核雄性不育两用系的可育株中分离到的雄性不育相关基因。本研究根据BcMF4基因的cDNA序列设计两对特异引物,从普通白菜花蕾cDNA中扩增出两个片断后连接至双元载体pBI121中,得到了RNAi(RNA interference)植物表达栽体pBI-B4R,并导入了农杆菌LBA4404菌株中;通过组织培养途径转化菜心(B．campestris ssp．chinensis var,parachinensis),72．2％的菜心转基因植株中45．8％的花粉为缩小而空瘪的畸形,而且这些植株的花粉离体萌发率降低至23．7％;Northern杂交显示,转基因植株的花粉畸形,是由于BcMF4基因片段的插入使BcMF4基因的表达受到了抑制。结果表明,采用RNAi技术下调了BcMF4基因的表达,导致了菜心转基因植株部分花粉的不育,证明BcMF4基因在普通白菜和菜心等白菜植物的花粉发育中起着重要作用。

油菜叶绿素b减少突变体Cr3529叶绿素生物合成的研究

利用吸收光谱和荧光光谱法测定了油菜叶绿素b减少突变体Cr3529子叶叶绿素生物合成途径中几种主要前体物质的含量.结果显示:突变体子叶中叶绿素生物合成第一个限速步骤的前体物质δ-氨基乙酰丙酸(ALA)含量与野生型油菜大致相同,饲喂ALA后的突变体及野生型油菜子叶中ALA含量均显著增加,但二者无显著差异;胆色素原含量在突变体中也未降低,而尿卟啉原Ⅲ含量仅为野生型的一半,粪卟啉原Ⅲ、原卟啉Ⅸ、镁原卟啉Ⅸ和原植基叶绿素的含量都明显低于野生型.结果证明,Cr3529突变体中叶绿素生物合成受阻于由胆色素原形成尿卟啉原Ⅲ的步骤,其叶绿素合成缺陷的机制和前体物质的累积与其它叶绿素b减少突变体明显不同.

田野菟丝子(Cuscuta campestris)寄生对薇甘菊(Mikania micrantha)入侵群落土壤微生物生物量和酶活性的影响

比较分析了广东省内伶仃岛薇甘菊未入侵群落、薇甘菊入侵群落、田野菟丝子刚寄生的薇甘菊入侵群落和田野菟丝子寄生3 a的薇甘菊入侵群落的土壤化学特性、微生物生物量碳氮磷及土壤酶活性的变化,旨在探讨薇甘菊入侵如何改变土壤特性及田野菟丝子的寄生如何改变薇甘菊入侵地土壤特性。薇甘菊入侵群落土壤的pH值(6.046)、有机碳(35.937 g.kg-1)、全氮(2.449 g.kg-1)、有机氮(2.383 g.kg-1)和氨态氮(0.051 g.kg-1)含量要显著地高于薇甘菊未入侵群落土壤(5.593,29.512g.kg-1,0.800 g.kg-1,0.722 g.kg-1,0.043 g.kg-1),而土壤硝态氮含量(0.015 g.kg-1)要显著地低于薇甘菊未入侵群落土壤(0.033 g.kg-1),土壤全磷和有效磷没有明显的差异;薇甘菊入侵群落土壤的微生物生物量碳、氮、磷、土壤酸性磷酸酶、脲酶和β-D-葡萄糖苷酶活性要显著地高于薇甘菊未入侵群落土壤。田野菟丝子寄生可以使薇甘菊入侵地的土壤pH值(5.634)、有机碳(27.225 g.kg-1)、全氮(1.836 g.kg-1)、有机氮(1.793 g.kg-1)和氨态氮(0.024 g.kg-1)含量显著性下降,对于全磷、有效磷和硝态氮则无明显影响;同时田野菟丝子寄生可以使土壤微生物生物量碳、氮、磷、土壤酸性磷酸酶、脲酶及β-D-葡萄糖苷酶活性显著下降,但改变后的土壤与未入侵地之间仍具有一定的差异。田野菟丝子寄生达3 a的薇甘菊入侵地的土壤总有机碳(35.719 g.kg-1)、全氮(2.356 g.kg-1)、有机氮(2.304 g.kg-1)和氨态氮(0.040 g.kg-1)含量相对于寄生早期显著增加,有机碳、全氮、有机氮等含量恢复到薇甘菊入侵地的水平,与未入侵地之间存在显著性差异;田野菟丝子寄生时间对土壤微生物生物量氮磷及土壤酸性磷酸酶和β-D-葡萄糖苷酶活性无显著性影响,但微生物生物量碳及脲酶活性显著升高,甚至超出薇甘菊入侵地。薇甘菊入侵可以改变土壤微生物生物量和酶活性,最终改变土壤化学特性,有利于其入侵;而田野菟丝子寄生可以打破土壤微生物生态系统的动态平衡,引起土壤微生物生物量和酶活性的改变,而最终又引起土壤化学特性的改变。此研究结果对于评价薇甘菊入侵的后果、田野菟丝子防治的可能机制及带来的后果具有重要的意义。

UV-B与AR胁迫下油菜光合及CAT活性的恢复过程(Ⅱ)

采用水培试验方法研究了油菜幼苗保护酶(CAT)及光合作用受紫外辐射(UV-B,280～320nm)与酸雨(AR)单一和复合胁迫后的恢复过程,考察了各项指标在胁迫后0～64h中的变化。结果显示,AR(pH3.5)对E%、CAT活性、Chl含量及En的影响是可恢复的,而对Pn造成了不可逆伤害;UV-B(0.45W·m-2)对E%、CAT活性、Chl含量、Pn及En的影响均可恢复;AR+UV-B对E%、CAT活性、Chl含量、En影响可恢复,而对Pn的影响无法恢复。试验结果还进一步说明,植物自身修复难易程度及恢复进程的快慢都与胁迫方式有关。

油菜光合作用及CAT对UV-B与AR胁迫的响应(Ⅰ)

采用水培试验方法研究了紫外辐射(UV-B,280～320nm)与酸雨(AR)单一和复合胁迫对油菜幼苗保护酶(CAT)及光合作用的影响。结果表明,单一胁迫与复合胁迫对油菜幼苗CAT活性及光合作用有不同程度的作用效果。在单一UV-B辐射(0.45W·m-2),单一酸雨(pH3.5)胁迫及复合胁迫下,质膜透性、MDA含量及CAT活性的增幅为8%、18.1%、26.0%,11.5%、21.6%、66.7%和28.8%、85.0%、113.8%;叶绿素含量、净光合速率、蒸腾速率、水分利用效率及气孔导度降幅为16.5%、48.6%、57.2%,28.6%、42.9%、78.6%,3.9%、12.4%、14.8%,27.8%、31.0%、74.5%和35.6%、38.2%、44.7%;而胞间CO2浓度的增幅为9.3%、10.6%、26.4%。结果显示,复合胁迫的抑制效应强于单一胁迫,即UV-B+AR>UV-B>AR,且对不同指标的影响不尽一致。

白菜型油菜双显性核不育896AB恢复系基因型的鉴定

以白菜型油菜双显性核不育896AB为材料,采用成对兄妹交和相应可育株自交,验证显性不育基因的遗传;用恢复系与全不育系测交,测交一代与临保系复交,验证显性恢复基因的抑制作用,并区分F_2代育性分离为3:1和13:3的遗传类型.经4个年度的研究认为,育性是由一对显性不育基因MSMS和一对显性可育基因RfRf互作控制,且显性可育基因能抑制不育基因的表达,使不育性恢复可育.

稀土对UV-B辐射伤害植物的影响:Ⅱ.Ce对UV-B胁迫下油菜幼苗保护酶系统影响

水培法研究了Ce对紫外辐射(UV-B, 280～320 nm)胁迫下油菜幼苗膜保护酶的影响. 实验表明, UV-B辐射(T1/0.15 W·m-2和T2/0.35 W·m-2)胁迫下, 油菜幼苗叶绿素含量减少, 质膜透性增大, SOD, CAT与POD酶活随时间进程先升后降(T1), 高剂量(T2)下POD持续升高, 3者响应UV-B辐射胁迫的敏感序列是SOD＞CAT＞POD. Ce对3种保护酶的调控作用, 减轻了UV-B辐射对其功能的损伤, 增强了保护酶清除自由基能力, 维持了质膜正常透性, 且对低剂量(T1)的防护效果优于高剂量(T2). 从而在防御系统层面实现了Ce对UV-B辐射伤害油菜幼苗的防护效应.