甘蓝型油菜小孢子离体诱变——紫外线对小孢子胚状体再生的影响

用紫外线对3个甘蓝型油菜品系的离体小孢子进行照射0-60s。未经照射的对照和已照射的小孢子于NLN培养基进行培养,诱导胚状体。结果表明,照射20s和60s对胚状体的再生没有影响,照射40s则有促进作用。     

具双价基因的甘蓝型油菜游离小孢子的遗传转化

以双低(低芥酶和低硫甙)甘蓝型油菜(Brassica napus)游离小孢子为受体，通过根癌农杆菌(Agrobacterium tumefaciens)介导，将几丁质酶基因和β-1，3-葡聚糖酶基因导入杂交油菜新品种油杂4号亲本恢复系和保持系中，成功地获得了转基因植株。在小孢子数与根癌农杆菌数接近时，100-200mg／L羧苄青霉素能有效地抑制菌的生长，而不影响小孢子胚胎形成。用预培养2d的小孢子与根癌农杆菌共培养10h后离心，重新悬浮小孢子于含上述浓度的羧苄青霉素的NLN培养基中培养，获得了16株抗卡那霉素的恢复系的单倍体植株。经PCR及PCR-Southern blot检测，有10株呈阳性，证明外源基因已整合到油菜基因组中。     

利用小孢子胚离体培养筛选抗除草剂的油菜

选用4个甘蓝型油菜（Brassica napus L．）F、（7039，7040，282和5102）为小孢子培养供体材料，分别对小孢子胚在含草甘膦和盖草能的培养基上进行培养，以筛选出抗草甘膦和抗盖草能的胚状体，进而获得再生植株。其中，基因型7039和7040用于草甘膦筛选，282和5102用于盖草能的筛选。选取子叶期胚状体，在含0．006％的草甘膦及0．01％和0．02％盖草能的MS-2培养基上培养2周，不抗草甘膦和小抗盖草能的胚状体2周内变褐死去，抗草甘膦和抗盖草能的胚状体转绿。随后转绿胚状体转移至正常MS-2培养基中继续培养，直至获得再生植株。再生植株喷施0．25％的草甘膦液表明均抗草甘膦；而对植株喷施0．05％的盖草能液时，0．02％筛选出的植株大部分为抗盖草能，而0．01％筛选出的植株却大部分死去，表明用0．02％浓度筛选抗盖草能植株的方法更有效。用170mg／L秋水仙碱直接处理再生植株20和30h，染色体加倍率分别达到了34％和52％。     

四倍体青花菜小孢子培养及胚胎发育途径研究

以同源四倍体青花菜为试材,进行了游离小孢子培养的胚胎诱导条件及胚胎发育途径的研究。结果表明:基因型是影响四倍体青花菜小孢子胚胎诱导率的重要因子,基因型A36的出胚率最高,达到28.5个·蕾-1;热激胁迫(32.5℃)诱导小孢子从配子体发育途径向胚胎发生途径转换,其小孢子胚胎发育途径为典型的单核期小孢子经对称分裂产生胚胎的途径(B途径);4℃低温预处理1d或2d结合32.5℃热激1 d后,对称分裂和多细胞团增加,供试基因型小孢子出胚率显著提高。本研究将有助于游离小孢子胚胎发生机理的研究,以及其他芸薹属小孢子胚胎体系的建立及优化。     

甘蓝型油菜越冬死苗与环境条件的关系及其防御措施

甘蓝型油菜越冬死苗除与初冬低温冻害有着密切的关系外，并与品种和栽培措施有关，选用科决策或冬性较强的品种和加强栽培管理措施，可减轻和预防甘蓝型油菜越冬死苗。     

甘蓝型油菜硼营养高效的遗传分析

利用土壤有效硼缺乏的田间小区试验 ,研究甘蓝型油菜硼营养高效品种和低效品种间 2个杂交二代群体及其回交一代硼营养性状的分离规律。结果发现 ,硼营养高效受一对主效基因控制。在缺硼胁迫下 ,硼高效性状与抽薹天数和生育天数之间紧密连锁 ,生育期 (抽薹天数和生育天数 )可作为甘蓝型油菜硼高效种质筛选及其杂种后代硼效率鉴定和选育的参考指标。     

硼对甘蓝型油菜蛋白质的影响

利用比色法和聚丙烯凝胶电泳法分析了不同硼营养状况下甘蓝型油菜的幼苗叶片及开花期花药内可溶性蛋白质含量及组分的变化。结果表明,缺硼或硼过量部会造成幼苗叶片及花药内蛋白质含量显著下降;从电泳图谱中也可看出,可溶性蛋白质的组分也发生了变化。不适当供硼(缺硼或硼过量)幼苗叶片及花药内所测的大部分蛋白质含量减少,有个别的蛋白质增加或缺失。     

低硼对甘蓝型黄籽油菜与黑籽油菜花期生理特性的影响

为探索甘蓝型黄籽油菜与黑籽油菜花期相关生理特性对低硼的响应差异,以2个具有相同遗传来源的黄籽(L1)和黑籽(L2)油菜基因型为材料,研究了田间条件下低硼处理对甘蓝型黄、黑籽油菜花期部分生理特性的影响。结果表明,低硼处理后黄籽油菜下部叶片中养分的再利用能力降低,而黑籽油菜仍有一定的养分可供利用;无论黄籽油菜还是黑籽油菜,低硼处理均使其花器官形态变小,干重下降,低硼对黄籽油菜雌雄蕊长度的影响大于黑籽;甘蓝型黄、黑籽油菜的花粉活力在低硼处理后明显降低,但黄籽油菜无活力花粉的比率明显高于黑籽油菜;低硼使甘蓝型油菜雄蕊和雌蕊中的可溶性糖、淀粉、游离脯氨酸以及可溶性蛋白质含量降低,黄籽油菜的降低幅度明显大于黑籽油菜。与相同遗传来源的黑籽油菜相比,甘蓝型黄籽油菜花期的相关生理特性更易受低硼的影响。     

小孢子培养获得松花型花椰菜DH再生植株

对5个松花型花椰菜(Brassica oleraceavar.botrytisL.)杂种一代的小孢子培养研究表明,小孢子胎胚发生主要依赖于基因型,庆农65天的每花蕾胚状体产量最高,平均达15.5个。松花菜的胚状体萌发率一般在30%左右,并有效地获得了大量的DH再生植株。冷激预处理能显著地影响花椰菜小孢子的胚胎发生,但供体材料间存在不同的结果。结球期和开花结角期再生植株的生育期与育性出现较大分离,可育且能正常结角的比例约占全部小孢子再生植株的50%以上,因而不再需要加倍处理。     

比较研究苏北沿海滩涂盐土上不同油菜品种生物学特征和产量构成

以实验室自育的5个油菜品种(系)及其他6份油菜品种(系)为试验材料,研究了这11份油菜品种(系)在轻(中)度盐化土中的越冬性状、生物学特性、产量构成特性,得出如下结论:南盐油系列品系(种)在盐土上植株生长势强,从基部开始分枝,分枝多;南盐油系列品种(系)茎粗与宁油16、三北98、秦油66和秦油9号差异不明显,但高于宝杂油1号和秦油33;各品种(系)株高差异不大;荚长与荚宽差异也不大。南盐油系列品系(种)产量显著高于对比的宝杂油1号、秦油66、秦油9和秦油33,一次与二次分枝数与菜籽产量的回归分析显示其R2均达极显著水平。     

青菜的原生质体培养

从青菜的下胚轴和根原生质体培养获得大量的愈伤组织并形成根。比较了影响青菜下胚轴和根原生质体分离的因素。培养基、培养方式、激素、椰子水、芸苔素等对培养效果有不同程度的影响。探讨了与青菜原生质体培养褐化有关的一些因素。进行了多种分化试验     

铝胁迫对油菜根系形态和生理指标的影响

为探讨油菜幼苗在不同浓度铝胁迫下的生理响应,以2个耐铝毒差异显著的油菜品种(耐铝品种R178和铝敏感品种S169)为试验材料,在营养液培养条件下,共设置0(对照)、50、100、150、200和 300 μmol·L^-1 6个铝(pH值4.5)处理,研究铝胁迫对油菜根系形态及生理指标的影响。结果表明,50 μmol·L^-1铝处理下R178各根系形态指标与对照相比均无显著差异,而S169根系各形态指标与对照均差异显著,其中总根尖数随着铝处理浓度的增加呈先升高后降低的趋势,其他根系指标均随着铝处理浓度的增加呈逐渐减少的趋势,300 μmol·L^-1铝处理下,2个品种的主根相对伸长率较对照分别减少65.7%和79.7%。随着铝处理浓度的增加,2个品种幼苗根系中可溶性蛋白和脯氨酸含量均呈先升高后降低的趋势,叶片中可溶性蛋白含量呈降低趋势,而脯氨酸含量则呈升高的趋势。铝胁迫导致2个品种幼苗根系和叶片的质膜透性显著增加,300 μmol·L^-1铝处理下R178和S169根系相对电解质外渗率较对照分别增加54%和59%,叶片中分别增加82%和90%,同一处理下R178根系和叶片电解质外渗率均小于S169。随着铝处理浓度的增加,2个品种根系中的超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性均呈先增加后降低的趋势;R178叶片中SOD活性呈增加趋势,而S169则呈先增加后降低的趋势,且在200 μmol·L^-1时达到最大值;2个品种叶片中的POD、CAT和抗坏血酸过氧化物酶(APX)活性均呈先增加后降低的趋势。综上,铝胁迫显著抑制了油菜根系生长,破坏了细胞质膜的完整性,油菜幼苗通过蛋白质和脯氨酸含量的积累以及保护酶活性的增高抵抗铝胁迫。本研究结果为耐铝油菜资源筛选和新品种选育提供了理论依据。     

甘蓝型油菜EMS诱变后代农艺性状观察及分子检测

利用不同浓度甲基磺酸乙酯(EMS)溶液处理甘蓝型油菜常规品种川油20和渝黄1号选系种子,结果表明:EMS对成苗率的抑制作用随着浓度的增加而增强,浓度1.0%为半致死量。M1中出现叶片卷曲、皱缩、黄化、双茎、扁茎和植株矮化等变异性状;M2中筛选到双茎、长角、多分枝等变异性状。对M2长角果等产量性状变异株进行考察,结果表明变异株角果长度(果身和果喙)高于对照,有效分枝数、有效角果、角果粒数和千粒重高于对照,一次分枝高度低于对照。对变异株进行SSR分析表明,变异株与对照在电泳条带上表现出多态性,进一步证明了变异的真实性。     

Efficiency of foliar selenium application on oilseed rape (Brassica napus L.) as influenced by rainfall and soil characteristics

The Czech Republic is characterized by a low Se soil content, resulting in Se deficiency in crops, humans, and animals. This study investigated the response of oilseed rape to foliar application of selenate solution in a microscale field experiment conducted at two locations differing in soil and climatic conditions but with comparable total Se contents. Sodium selenate (Na₂SeO₄) was applied at two rates (25 and 50 g Se ha^?1). The potential effect of Se application on the uptake of essential elements was also evaluated. The foliar Se application resulted in an effective stepwise increase in the Se contents of all the plant components studied (leaves > stems > roots > siliques ~ seeds), as expected. No significant influence of Se fortification on the other investigated macro- and microelements was observed. However, the soil and climatic conditions influenced the Se uptake, such that a higher Se content was observed in plants grown in the most acidic location (Cambisol soil) that had a higher oxidizable carbon content and higher average annual rainfall compared to the less acidic location (Luvisol soil). These observations indicated the necessity to optimize the Se application for the particular soil and climatic conditions to achieve a maximum biofortification effect.     

辐照诱导的新雄性不育系过氧化物酶和脂酶同工酶分析

采用聚丙烯酰胺凝胶垂直板电泳法,对甘蓝型油菜细胞质雄性不育系Polima、Shan2A、Ogu和辐照处理的新雄性不育系Xin1和Xin2及其保持系Shan2B的种子进行了脂酶和过氧化物酶分析,结果表明:5个细胞质雄性不育系比保持系Shan2B多1条迁移率为0.45的脂酶(EST)谱带;Xin1、Shan2A和Polima各有4条过氧化物酶(POD)谱带,保持系缺少1条迁移率为0.61的POD谱带,推测这一同工酶的表达可能与雄性不育相关。     

Impact of arbuscular mycorrhizal fungi on the growth and expression of gene encoding stress protein – metallothionein BnMT2 in the non‐host crop Brassica napus L.

Arbuscular mycorrhizal (AM) fungi are an important component of the soil biota in most agroecosystems, and their association can directly or indirectly affect the diversity of soil microorganisms, nutrient cycling, and growth of host plants. Since not all crops are symbiotic, we hypothesized that the presence of AM fungi can: (1) inhibit the growth of non‐host plants by resulting in biotic stress, or (2) promote their growth indirectly by increased nutrient mobilization. These hypotheses were tested in the present study on the non‐mycorrhizal crop canola (Brassica napus L.) in the presence and absence of other autochthonous soil microorganisms. The soil was inoculated with a mixture of AM fungi (Acaulospora longula, Glomus geosporum, G. mosseae, Scutellospora calospora) and as a control, a non‐inoculated soil was used. The impact of inoculation on plant growth (biomass production, nutrient concentrations) and expression of the stress protein metallothionein gene BnMT2 was investigated in the shoots. B. napus L. did not form mycorrhizal associations on its roots, but its growth was promoted after inoculation with AM fungi. In the soil with autochthonic microorganisms, growth inhibition after inoculation was observed compared to the control. The concentrations of N, P, K, and S in the shoot were always significantly increased after inoculation with AM fungi. However, this was partly combined with reduced growth and thereby decreased total uptake of nutrients. Expression of BnMT2 in the leaves was increased after inoculation with AM spores at the soil devoid of indigenous microorganisms, but decreased in their presence. The expression of stress proteins (BnMT2) significantly increased with increasing length and biomass of shoots. In conclusion, the inhibition of the non‐host plant B. napus L. following inoculation with AM fungi was confirmed, however, only in combination with autochthonous microorganisms. Growth promotion of B. napus L. in the presence of AM fungi in the absence of autochthonous soil microorganisms suggest that plant growth depression in the presence of AM fungi was based on interactive effects of AM fungi with the autochthonous microorganisms in the soil rather than on a direct impact of the AM fungi.     

Regulatory effects of sulfur on oilseed rape (Brassica napus L.) response to selenite

Selenite is a form of selenium (Se) commonly found in Se-excessive soils. To regulate the Se content in plants in high-Se areas, a potted soil experiment was performed on oilseed rape (Brassica napus L.) to evaluate the effects of varied amounts of sulfur (S) on the biomass, accumulation and distribution of Se in B. napus under the conditions of different amounts of Se in the soil. The results showed that the seedlings of B. napus were more sensitive to Se than the mature plants were. The addition of S significantly alleviated the growth inhibition in seedlings and facilitated the growth of mature plants under higher Se (15 mg kg^?1) conditions. S treatment significantly decreased soil pH within the range of 0.22–0.60. An appropriate moderate amount (150 mg kg^?1) of S exerted the strongest inhibition on Se concentration and accumulation in B. napus at the seedling stage, but a higher amount (300 mg kg^?1) of S led to a more significant decrease in the mature plants under higher Se conditions, with the maximum reduction in various parts of B. napus reaching 51.3–60.9% and 42.5–53.4%, respectively. The application of S only affected the uptake of Se, and not the translocation of Se; the accumulation of Se in B. napus follows the sequence of pod ≈ stem > rapeseed > root, and the distribution ratio is approximately 1.00:0.97:0.69:0.49. Overall, the application of S alleviated the inhibitory effect on growth caused by excessive Se by reducing the Se concentration in B. napus and facilitating its growth, suggesting that S treatment is a suitable and highly cost-effective method to regulate the content of Se in B. napus.     

喷洒氯化钾对油菜植株硝酸盐再利用的影响

从植物体本身着手研究植物氮素高效利用的机理与途径,这是近几年来植物氮素营养研究的热点之一,而调动植物液泡中的硝酸盐使之得到更高程度的再利用,这是提高植物氮素利用效率和降低植物体内硝酸盐含量的途径之一。试验采用土培试验,研究叶面喷施质量分数为1%的氯化钾溶液对油菜角果初期和收获期植株硝酸盐累积再利用的影响,以喷施质量分数为1.17%的硫酸钾溶液(要求氯化钾和硫酸钾溶液中钾离子的物质的量浓度相同)和清水作为对照,在开花后期开始喷施,连续隔日喷洒15天,喷施部位在茎和叶,重点喷洒叶的背面,尽量不喷施至叶柄,于角果初期和收获期采样。结果表明,喷施氯化钾溶液能显著降低油菜角果初期和收获期茎、叶、根硝酸盐含量和植株硝酸盐总量,促进硝酸盐再利用;可以增强角果初期叶片硝酸还原酶的活性;能增加收获期干物质累积量,但不影响角果初期干物质累积量。