以叠抱类型和舒心类型四倍体大白菜为试材,采用实时荧光定量(qRT-PCR)方法对中脱落酸相关基因Bra018800和生长素相关基因Bra018750进行表达量分析,并利用酶联免疫法对其内源激素进行测定,研究了四倍体大白菜叶球形成内源激素变化及相关基因表达变化,以期为后继研究四倍体大白菜不同叶球类型形成的机理提供参考依据.结果 表明:Bra018800和Bra018750基因分别在叠抱类型四倍体大白菜的包心期和莲座期表达量达到最高,比舒心类型同时期表达量高2.56～2.80倍和5.18～6.59倍.内源激素测定结果表明,脱落酸(ABA)和生长素(IAA)2种激素在叠抱和舒心四倍体大白菜2类材料中表现为差异显著.其中ABA在叠抱类型四倍体大白菜的包心期含量达最高,比舒心类型四倍体大白菜同一时期的含量高2.05～2.12倍,与脱落酸相关基因Bra018800在2种类型材料中的表达趋势相一致;在叠抱类型四倍体大白菜莲座期IAA含量显著高于舒心类型同时期的2.58～2.60倍,与生长素相关基因Bra018750在2种类型材料中的表达趋势相一致.综上所述,Bra018750和Bra0188002个基因调控内源激素IAA和ABA的合成,分别在四倍体大白菜包心期和莲座期对叶球类型的形成起着关键性作用.

枣庄学院生物科学专业发展历史悠久,2018年被山东省教育厅确定为山东省公费师范招生专业,2019年被教育部列为山东省师范专业认证试点专业.为了响应国家号召,贯彻教育部规定,同时为培养"四有"生物科学师范专业优秀毕业生,枣庄学院生命科学学院通过综合分析内部质量监控和外部评价的结果,以追求卓越的质量文化建设,推动师范生培养质量持续改进.

植物科学与技术专业为农业高校传统专业.近几年,随着枣庄学院向应用型大学转型发展的要求,对植物科学与技术专业建设提出了新的要求,专业加强实践教学,培养富有创新精神和开拓能力,适于区域农业发展的应用型高级技术人才.文章旨在探讨植物科学与技术专业的建设,以适应鲁南地区经济发展的需要.

目前国内外对二倍体大白菜雄性不育系的创制已进行了深入的研究,利用雄性不育系技术育种已相对成熟,而四倍体大白菜育种仍以突破稔性、提高结籽率的自交系选育为主,四倍体雄性不育系的研究尚无相关报道.本研究通过从二倍体大白菜细胞质雄性不育系中筛选含新型甘蓝型油菜CMS基因的资源(CMS后36高)为不育源,依靠人工秋水仙素加倍,经倍性鉴定筛选和高代稳定四倍体自交系D571、D574连续多代回交转育,成功获得了CMS-D571、CMS-D574两个四倍体大白菜细胞质雄性不育系,该不育系具有不育度达100％、结籽率高、遗传稳定的特点.该研究证实细胞质雄性不育性也可在四倍体大白菜育种上应用,可以作为创制四倍体大白菜细胞质雄性不育系的方法,为四倍体大白菜开展雄性不育系育种奠定了物质基础.

目的 优化表面活性剂-超声波协同提取石榴瓤总黄酮工艺,并考察其动力学、热力学行为.方法 在单因素试验基础上,以总黄酮得率为评价指标,十二烷基硫酸钠(SDS)用量、乙醇体积分数、超声温度为影响因素,中心组合设计优化提取工艺.然后,考察提取过程中的动力学、热力学行为.结果 最佳条件为SDS用量0.84％,乙醇体积分数61％,超声温度53℃,总黄酮得率1.736％.加入表面活性剂(SDS)后,提取速率常数(k)、表面扩散系数(Ds)显著增加,提取为吸热熵增加的自发过程.结论 该方法稳定可靠,可用于表面活性剂-超声波协同提取石榴瓤总黄酮.

Based on the investigation of Zaozhuang City in Shandong Province,the present situation of rice production in Zaozhuang was analyzed from the aspects of natural ecological conditions,varieties planted,cultivation techniques,pests and diseases.Existing problems were pointed out in aspects of cultivars,cost effectiveness of planting,irrigation and drainage facilities,production techniques,industrialization level,local governmen-tal management,and scientific research investment.Targeted countermeasures to promote the development of rice industry of Zaozhuang City were also proposed.

为研究茶多酚对肉仔鸡生产性能及抗氧化能力的影响,试验选用了体重相近、健康的1日龄AA肉仔鸡(公)128只,随机分成4组,每组设4个重复,每个重复8只鸡.试验期为42 d.在各组基础饲料中分别添加0、40、80、120 mg/kg的茶多酚.结果表明:21日龄,120mg/kg茶多酚组的平均日采食量(ADFI)和平均日增重(ADG)分别显著高于对照组18.64％(P ＜0.05)、22.86％ (P ＜0.05),与对照组相比,120mg/kg茶多酚组的料肉比(F/G)则显著降低了3.42％(P＜ 0.05).而在42日龄时,与对照组相比,茶多酚各添加组的ADFI(P＜ 0.05)和ADG(P＜ 0.05)均显著增加,茶多酚80mg/kg与120mg/kg组的料肉比均显著降低(P＜0.05).21日龄时,与对照组相比,80、120mg/kg茶多酚组肉仔鸡血清中谷胱甘肤过氧化物酶(GSH-Px)、过氧化氢酶(CAT)和超氧化物歧化酶(SOD)的活力均显著上升(P＜0.05);而在42日龄时,80、120mg/kg茶多酚组肉仔鸡血清中GSH-Px和SOD的活力均显著上升(P＜0.05),但CAT活力无显著性变化(P＞0.05).在21日龄时,与对照组相比,各试验组之间肉鸡血清的丙二醛.(MDA)含量显著下降(P＜0.05).而在42日龄时,与对照组相比,茶多酚80、120 mg/kg组肉仔鸡血清中的MDA含量分别显著降低了14.88％、17.79％(P＜ 0.05).与对照组相比,茶多酚80、120mg/kg组的肉仔鸡胸肌GPx4表达水平显著上调.综上,茶多酚能促进肉仔鸡的生长,提高血清抗氧化物酶GSH-PX、SOD和CAT的活性,降低肉鸡血清中的MDA含量,显著提高肉鸡胸肌GPx4基因m RNA的表达量.

The ultrasonic-assisted extraction conditions of total flavonoids from pomegranate root bark were studied.Single factor test and Box-Behnken experimental design were used to optimize the extraction process employing total flavonoids yield as the response,and ethanol concentration,solid/liquid ratio,ultrasonic power and ultrasonic time were used as independent variables,and the nitrite scavenging activity and nitrosamine synthesis inhibiting activity were tested.Results showed that under the following conditions:Ethanol concentration of 63％,solid/liquid ratio of 1∶22 g/mL,ultrasonic power of 270 W and ultrasonic time of 34 min,the yield of total flavonoids was 2.81％,close to predicted value.The total flavonoids from pomegranate root bark had a strong nitrite scavenging activity and nitrosamine synthesis inhibiting activity.At the concentration of 3.6 μg/mL,the total flavonoids exhibited a maximum sodium nitrite scavenging rate of 64.5％ and the effect of IC50 =2.408 μg/mL.At the same time,the total flavonoids exhibited a maximum nitrosamine synthesis inhibiting rate of 71.9％ at the effect of IC50 =2.345 μg/mL.There was a dose-response relationship between the total flavonoids from pomegranate root bark and the nitrite scavenging activity and nitrosamine synthesis inhibiting activity in the range of 0.12～ 3.6 μg/mL of concentration.This method could provide a scientific basis for the extraction and application of total tlavonoids from pomegranate root bark.

The cold resistance of 24 pomegranate cultivars was evaluated by the changes of physiological and biochemical indexes under low temperature stress with one-year-old dormant shoots as materials.Adopting simulated low temperature (room temperature,0,-4,-8,-12,-16,-20℃) for low temperature treatment,the relative conductivity (REC),semilethal temperature (LT50) and proline (Pro) and malondialdehyde (MDA) contents were determined and the cold resistance of 24 pomegranate cultivars was evaluated comprehensively by membership function method.The results showed that the REC,Pro and MDA contents of one-year-old shoots of 24 pomegranate cultivars showed an increasing trend with the decreasing of stress temperature.The LT50 of Yicheng Kanghan 1 was the lowest,which was-14.32℃ and showed the best cold resistance;the LT50 of Tunisi soft-seed pomegranate was the highest,which was-7.95 ℃ and showed the worst cold resistance.Through the membership function method for comprehensive evaluation of cold-resistance,we could concluded that 8 cultivars were classified into the grade Ⅱ with cold resistance,15 cultivars were classified into the grade Ⅲ with moderate resistance and 1 cultivar belonged to the grade Ⅳ with low resistance.

The ionic liquid based ultrasonic-assisted extraction conditions of proanthocyanidins fron pomegranate seeds were studied.Single factor test and Box-Behnken experimental design were used to optimize the extraction process employing proanthocyanidins yield as the response,and extraction time,extraction temperature,ultrasonic power and concentration of ionic liquid were used as independent variables.The kinetics and thermodynamics of extraction process was studied basing the optimized extraction conditions.Results showed that the optimal extraction conditions for proanthocyanidins were determined as follows:extraction time of 32 min,extraction temperature of 52 ℃,ultrasonic power of 300 W,and concentration of ionic liquid of 1.15 mol/L.Under these conditions,the yield of proanthocyanidins was 1.718％.Kinetic parameters showed that extraction rate constant k value was 0.313 min-1,surface diffusion coefficient Ds value was 7.936 × 10-5 cm2/min while those of ethanol control group were 0.063 min-1 and 1.597 × 10-5 cm2/min,respectively,and the extraction rate was significantly improved.Thermodynamics parameters showed that entropy ΔH was 248.51 kJ/mol,enthalpy ΔS was 813.69 J/ (mol· K),the values were all greater than zero,indicating that the ionic liquid extraction process was an endothermic process of increasing entropy,at the same time,the free energy ΔG was-18.254 kJ/mol,less than zero,indicating that the ionic liquid extraction process was a spontaneous process.Both the optimized ionic liquid extraction technique and dynamics/thermodynamics analysis can be used for industrial scale-up and optimal control of proanthocyanidins extraction from pomegranate seeds.

This study aimed to optimize the ultrasonic-assisted cellulase-based extraction of polyphenols from pomegranate fruitlets. The factors with a significant influence on the yield of polyphenols were selected using a combination of one-factor-at-a-time method and Plackett-Burman (PB) design for the optimization experiments which were carried out using Box-Behnken design combined with response surface methodology. The anti-α-glucosidase activity and kinetics characteristics of the polyphenols were determined by using α-glucosidase inhibition model in vitro. Results showed that under the following conditions: ultrasonic power, 318 W; enzyme dosage, 23 U/mL; hydrolysis temperature, 46 ℃; and hydrolysis time, 2.7 h, the average yield of polyphenols from pomegranate fruitlets was 11.65%, close to the predicted value. The polyphenols had a strong anti-α-glucosidase activity. The percentage inhibition of α-glucosidase by the polyphenols at a concentration of 1.20 mg/mL reached 70.3% and the half inhibitory concentration (IC50) was 0.747 mg/mL. There was a positive relationship between the anti-α-glucosidase activity and polyphenol concentration in the range of 0.24–1.20 mg/mL. The inhibition was reversible and non-competitive.

The effect of different allicin supplemental levels in Arbor Acres (AA) broiler feed on lipid metabolism and related genes expression were studied .A total of 96 ,1 d old AA broilers were assigned to 4 diet treatments (0 ,200 ,400 ,and 600 mg/kg) with 4 replications of 6 birds for 42 d .The results showed that increasing the levels of allicin significantly decreased percentage of liver fat and percentage of abdominal fat (P<0 .05) ,but no significant differences in thicknesses of subcutanceous adipose and intermuscular adipose tissue were observed among the treatments (P>0 .05) .Compared with the control group , serum triglyceride (TG) ,total cholesterol (TC) and low density lipoprotein(LDL-C)concentration were significantly decreased in 400 and 600 mg/kg allicin groups (P<0 .05) ,but the level of high density lipoprotein (HDL-C)was significantly increased (P<0 .05) .There was a trend of decline of the level of nonesterified fatty acid (NEFA) with the increase of the concentration of allicin among the treatments ,but the difference was not significant ( P> 0 .05 ) .In liver of AA broilers ,supplementing 400 mg/kg and 600 mg/kg allicin can decrease ACC and FAS mRNA expression (P< 0 .05);and the expression of LPL in 600 mg/kg allicin group was significantly higher than the control group (P<0 .05) .In abdominal fat ,the level of ACC was declinedsignificantly in 600 mg/kg allicin group(P<0 .05) compared with the control group ,and the expression of LPL was increased (P<0 .05) .However ,allicin had no influence on the expression of FAS mRNA in abdominal fat .

AIM To optimize the subcritical aqueous extraction for polysaccharides from the leaves of Punica granatum L.and to evaluate the in vitro antioxidant activity.METHODS With reaction pressure,solid-liquid ratio,extraction time and extraction temperature as influencing factors,yield of polysaccharides as an evalution index,the extraction was optimized by Box-Behnken method on the basis of single factor test.Then the scavenging effects of polysaccharides on hydroxyl free radical,superoxide anion and DPPH free radical were detected.RESULTS The optimal conditions were determined to be 5 MPa for reaction pressure,1 ∶ 27 for solid-liquid ratio,11 min for extraction time,and 155 ℃ for extraction temperature,the yield of polysaccharides was 1.809％.There was a dose-effect relationship between scavenging rate and polysaccharides' concentration.0.1 mg/mL Polysaccharides displayed the strongest scavenging effects on hydroxyl free radical,superoxide anion and DPPH free radical with the clearance rates of 57.36％,70.51％ and 58.02％,respectively.CONCLUSION This stable and reliable method can be used for the subcritical aqueous extraction for polysaccharides from P.granatum leaves with obvious in vitro antioxidant activity.

[Objective] The objective of this study is to reveal the karyotypic polymorphism ofAegilops speltoides (Aegilops short for Ae.hereafter) and the karyotypic difference between common wheat and Ae.speltoides via the establishment of FISH karyotype of Ae.speltoides.[Method] Multicolor fluorescence in siru hybridization (mc-FISH) was employed to detect the distribution of Oligo-pSc119.2 and Oligo-pTa535 in chromosomes of Ae.speltoides;centromere-specific oligonucleotide CCS1 wasused to identify the location of centromeres on chromosomes;FISH karyotype comparison was conducted to show the karyotypic differences between Ae.speltoides and wheat.[Result] In wheat,oligo-pTa535 signals were observed mainly on chromosomes in A and D genomes,and only very sporadic signals were found in B genome.However,oligo-pTa535 signals were absent in Ae.speltoides of five accessions.Oligo-pSc1l9.2,compared to a little distribution in A and D genomes,shined plentiful fluorescence in the whole genome B in wheat,and especially in S genomes in Ae.speltoides of five accessions used in this study.Different pairs of chromosomes in wheat could be distinguished from each other according to the distribution of Oligo-pSc119.2 and Oligo-pTa535 on chromosomes of wheat.FISH patterns produced by Oligo-.pSc119.2 in wheat showed similarity among wheat materials whether of different ploidy or of different varieties of same ploidy,and that in Ae.speltoides varied depending on accessions.Even homologous chromosomes in one cell in Ae.speltoides exhibited differences in FISH pattern.Oligo-pSc119.2 FISH patterns of five accessions each showed obvious differences from that of B genomes in wheat.Four of five Ae.speltoides accessions possess six pairs of metacentric chromosomes except for homologous pairs 4S of submetacentric chromosomes,which were involved in the karyotype formula 2n =14 =12m + 2sm,and the rest one,PI542238,however,houses seven pairs of metacentric chromosomes which resulted in the karyotype formula 2n =14 =14m.[Conclusion] Chromosomes of Ae.speltoides house rich repetitive DNA sequences highly homologous to pSc119.2 and lack that highly homologous to pTa53.The distribution ofpSc119.2 on chromosomes ofAe.speltoides showed differences between accessions,between plants of one accession and even between homologous chromosomes in one plant.FISH pattems produced by Oligo-pSc119.2 on Ae.speltoides chromosomes exhibited significant difference from that on chromosomes of B genomes in wheat.FISH analysis,using Oligo-pSc119.2 and Oligo-pTa535 as probes,not only could differentiate the chromosomes in wheat from that in Ae.speltoides,but also could dishtinguish the chromosomes from each other whether in wheat or in Ae.speltoides.

目的 优化复合酶(纤维素酶-果胶酶)提取石榴皮中原花青素的工艺,并建立其动力学模型.方法 以原花青素得率为评价指标,Plackett-Burman设计筛选影响因素,Box-Behnken设计对加酶量、酶解温度和酶解时间进行优化.建立4种提取工艺的动力学模型(Allometric1、Expdec1、Logistic、Sweibull1),通过精准因子(Af)、偏差因子(Bf)、根平均方差(RMSE)、决定系数(R2)分析模型的适用性.结果 最佳条件为加酶量1.5％、酶解温度48℃、酶解时间270 min,原花青素得率为1.88％.Logistic模型和Sweibull1模型拟合精度均较高,并且后者在最佳酶解温度(48℃)下预测效果更好.结论 所建立的优化条件和动力学模型均可用于石榴皮中原花青素提取工艺的工程放大和优化控制.

目的:优化石榴叶总黄酮的提取工艺,并对其体外抗氧化活性进行评价,为该部位的开发提供参考.方法:在单因素试验基础上,选择乙醇体积分数、料液比、超声功率、超声时间为考察因素,石榴叶总黄酮提取率为评价指标,根据Box-Behnken设计原理,采用响应面法优化石榴叶总黄酮的提取工艺.通过DPPH和·OH的清除能力考察石榴叶总黄酮的体外抗氧化活性.结果:石榴叶总黄酮最佳超声提取工艺为乙醇体积分数59.8％,料液比1∶16.3,超声功率221W,提取时间33.45 min;石榴叶总黄酮提取率0.433％,与理论值偏差很小.当样品质量浓度为0.6 ～1.0g·L-1时,对DPPH和·OH清除率较高,与维生素C接近.结论:优选的提取工艺稳定可行、提取率高,石榴叶总黄酮抗氧化性能较强,为该部位的工业化生产提供参考.

以石榴皮多糖与亚硒酸钠为原料制备石榴皮多糖硒酸酯,以产物硒含量和收率为指标,原料配比、硝酸体积分数、反应温度和反应时间为单因素,通过单因素试验和Box-Behnken试验设计方法对工艺条件进行优化,并对产物结构进行分析。结果显示:石榴皮多糖硒酸酯最佳制备工艺条件为原料配比1:1.0（g/g）、硝酸体积分数0.61%、反应温度77.0℃、反应时间11.3 h,在该工艺条件下,制得石榴皮多糖硒酸酯中硒含量为4.48 mg/g,产物收率为41.87%;紫外光谱、红外光谱和热重分析充分证实石榴皮多糖硒酸酯中含有Se=O键、Se—O键和Se—C键。优化后的石榴皮多糖硒酸酯制备工艺条件实现了石榴皮多糖的硒化,为石榴皮资源的充分开发利用提供良好条件。

In this study, we optimized the conditions for enzymatic extraction of polysaccharides from pomegranate seeds with a mixed enzyme preparation consisting of pectinase, cellulase and mannase. The optimization of the mixing ratio of the three enzymes was performed by the combined use of one-factor-at-a-time method andD-optimal mixture design. Thereafter, the enzymatic extraction conditions were optimized by response surface methodology using Box-Behnken design. The yield of polysaccharides was taken as response for both optimizations. It was shown that the optimal combination of enzymes were 24.2% pectinase, 67.2% cellulase and 8.6% mannose and that the optimal extraction conditions were determined as follows: liquid/solid ratio, 20:1 (mL/g); initial pH, 4.5; hydrolysis temperature, 48.5℃; and hydrolysis time, 287 min. Under these conditions, the yield of polysaccharides was 2.83%.

Objective] Powdery mildew is one of the most devastating diseases of wheat. It is widely accepted that the most economic, efficient and safest way to control powdery mildew is breeding and planting powdery mildew resistant cultivars.Aegilops ventricosa andAegilops cylindrica, which possess many favorable characters and good qualities such as resistance to diseases, tolerance to environmental stresses, are close-related relatives of wheat. The objective of this study was to develop novel powdery mildew resistant germplasm line via wide hybridization between common wheat andAegilops ventricosa and/orAegilops cylindrica for genetic improvement.[Method] Improved fuchsin squash and seed set examination were used to determine the cytogenetic stability and fertility of TA002. Acid polyacrylamide gel electrophoresis (A-PAGE) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) were used to analyze the subunit composition of gliadin, high and low molecular weight glutenin (HMW-GS and LMW-GS) respectively. Genomicin situ hybridization (GISH), multicolor genomicin situ hybridization (mc-GISH), multicolor fluorescencein situhybridization (mc-FISH) and molecular markers were employed to detect the genetic nature of TA002. Respective inoculation of powdery mildew isolates was performed to detect the reaction of TA002 to powdery mildew.[Result] TA002 (2n = 42) showed equivalent seed set to that of common wheat. Both TA002 and its hybrid F1 with common wheat Mingxian169 housed 21 bivalents in the pollen mother cells observed at metaphase I (PMCs MI) and took on chromosomal segregation of 21 to 21 in pollen mother cells examined at anaphase I (PMCs AI). Contrast to wheat parent Yannong15 highly susceptible to powdery mildew, TA002, similar to SDAU18 and its parentsAegilops ventricosa andAegilops cylindrica, was highly resistant to powdery mildew. Immunization and resistant-susceptible segregation of 3﹕1 was observed respectively in F1and F2 of the crosses between TA002 and common wheats susceptible to powdery mildew. The analysis of seed storage proteins of TA002 and its parents revealed that TA002 not only had new glutenin and gliadin subunits specific to SDAU18, but also possessed a novel gliadin subunit derived from its neither parents. When probed respectively by genomic DNA ofAegilops uniaristata andAegilops caudata in root tip cell chromosomes of TA002, no probe signals were detected. Multicolor genomicin situ hybridization (mc-GISH) and multicolor fluorescencein situ hybridization (mc-FISH) indicated that TA002 had three complete genomes (A, B and D) although five pairs of chromosomes 4A, 5A,6B, 7B and 5D in it showed significantly different FISH pattern from that of corresponding ones in Yannong15. Mc-GISH was carried out simultaneously using genomic DNA of Triticum uratu and that ofAegilops tauschii labeled with different fluoresceins as probes, and with genomic DNA ofAegilops speltoides as blocker. Mc-FISH was performed with two oligonucleotidespSc119.2 and pTa-535 fluorescing differently. Molecular marker analysis revealed TA002 housed alien genetic materials not only fromAegilops ventricosa but also fromAegilops cylindrica.[Conclusion]TA002, derived fromAegilops ventricosa orAegilops cylindrica with high powdery mildew resistance controlled by a single dominant gene, was found as a novel wheat-Aegilops introgression line of cytogenetic stability and good fertility. In addition, TA002 possessed novel seed storage protein subunits either specific toAegilops ventricosa -Aegilops cylindricaamphiploid or in neither parents.