小麦单细胞培养的研究──再生植株后代主要农艺性状变异及遗传稳定性

试验采用东北农业大学小麦生理和生物技术实验室获得的以普通小麦７７４２再生植株的２５个单细胞无性系为试验材料，对无性系后代株高和熟期等主要农艺性状进行分析．结果表明，株高和熟期等均发生明显变化，株高趋于交接，而且分离出明显接于对照（７７４２）的株系；熟期趋于提早，提早幅度大（５～８ｄ），多数株系均与矮秆结合．株高变矮和熟期提早在早期世代都可以稳定遗传．无性系后代茎秆、颖壳颜色及穗型等形态性状也发生变化，并可稳定遗传．表明小麦体细胞无性系变异是小麦矮秆早熟品种改良的有效途径．     

水稻离体培养及其无性系变异的研究

近30年来,水稻离体培养及其体细胞无性系变异的研究取得了很大的进展,已成为水稻品种改良中提高变异频率、增加变异类型、加快育种进程、提高育种效率等的一个重要手段.本文较全面地综述了有关资料,并提出一些粗浅看法.     

Studies on Immature Embryo Culture In vitro in Wheat： Genetic stability analysis of somatic variation in main agronomic chara

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The Application of Somaclonal Variation in Early Maturity,High Yield and High Quality Improvement in Wheat

Yield characters,maturity and grain protein content of somaclones derived both form immature embryo of cultivar 77(2)-Spring and single-cell culture of cultivar NE7742 in vitro were studied and the wide variatioin was found.Soaclones with maturity 8 days earlier than or the same as CK NE 7742(high yield,early maturity and high quality),combining with high quality(grain protein content 15.5%-18%)and high yield(the same as 7724 or higher)have been found and selected and now multiplied for 8 generations.The results of cultivar comparison trial in 1995 showed that several somaclones(the yields were significantly higher than CK DN 120) could be used directly in wheat production.The studies confirmed that somaclonal variation is one of the effective ways for early maturity,high-yielding and high-quality improvement in wheat.     

不同熟期冬小麦品种子粒蛋白质含量的遗传研究

采用Griffing方法Ⅰ ,利用 6× 6完全双列杂交 ,对不同熟期的 6个冬小麦品种 (系 )的子粒蛋白质的遗传机制进行了研究。结果表明 :不同品种子粒蛋白质含量的GCA效应、SCA效应和R效应均达极显著水平 ,其均方比为5 6 0 7∶1 383∶1。子粒蛋白质的遗传以加性基因效应占绝对优势。但反交效应和非加性基因效应也不容忽视。加性基因效应对后代蛋白质含量的提高因亲本而异 ,在该性状的遗传改良中存在着母体效应 ;蛋白质含量的广义遗传力为 90 95 % ,狭义遗传力为 5 4 99% ;各亲本的子粒蛋白质含量对后代传递的整齐度以河农 2 5 5 2为好     

水稻细胞悬浮系及其单细胞培养的研究

以水稻的不同遗传型材料的幼穗为外植体诱导愈伤组织,通过继代培养,最终筛选出了来自于品系“１４５”的胚性愈伤组织．在附加１．５～２．０ｍｇ／ｌ２,４—Ｄ的ＮＭＢ固体培养基上继代７次后,转入同种液体培养基上进行悬浮继代培养,由此形成的悬浮系是由单细胞和小细胞团组成的．倒置显微镜下几乎观察不到管形细胞,细胞的成活率达９０％以上．未发现酵母抽提物对细胞悬浮培养具有促进作用．在液体培养基中附加６００ｍｇ／ｌ的肌醇能够大大提高愈伤组织的分散程度,从中分离出的单细胞经初步培养再生了愈伤组织．     

小麦单细胞培养的研究——再生植株后代花粉母细胞染色体行为与花粉粒育性

在单细胞培养再生植株后代农艺性状和产量性状研究基础上 ,进一步对再生植株后代的花粉母细胞染色体行为和花粉粒育性进行研究。结果表明 ,花粉母细胞染色体行为发生异常 ,出现落后染色体、环形染色体、染色体桥、断片和粘连以及花粉母细胞仅发生核裂、三极纺锤体、多分体等 ,其中多以落后染色体花粉母细胞出现的频率为最高 ,早期世代为 8.0 3 %～ 51.11% ,随着繁殖世代的增加 ,这种异常的比率逐渐降低 ,到高世代 ( SCC6代 )仅为 0 .50 %～ 9.80 %。花粉粒育性分析结果表明 ,尽管早期世代花粉母细胞发生异常的比率较大 ,但各株系的可育花粉粒比例都在 60 %以上 ,并且均表现为正常结实 ,未发现不育现象。随着世代增加 ,育性也在不断改善 ,高世代绝大多数株系可育花粉粒的比例都在 98%以上。再生植株花粉粒的育性与花粉母细胞染色体变化密切有关     

优质冬小麦子粒灌浆过程中蛋白质和粒重积累动态的研究

选用8个加工品质优良的冬小麦推广品种。分析了子粒灌浆过程中蛋白质和粒重的积累动态。研究表明,优质小麦在子粒灌浆过程中N素的变化动态及累积趋势与非优质小麦品种基本相同。呈高,低,高凹型曲线。灌浆初期,子粒具有较高的蛋白质含量。随着子粒的增大。蛋白质含量逐渐降低。到开花后21d降到最低,之后又开始缓慢回升;粒重在灌浆前期增长迅速,后期增长缓慢;就群体而言,蛋白质积累与干物质积累密切相关,子粒产量和蛋白质产量呈同步增加,产量越高,蛋白质产量也越高。     

氮素供应对小麦子粒蛋白质组分及积累动态的影响

在施纯氮总量２４０ ｋｇ·ｈｍ － ２条件下,研究了等量氮肥不同基追比例和不同施肥时期对小麦子粒蛋白质组分及积累动态的影响,结果表明,氮肥不同基追比例各处理的醇溶蛋白和麦谷蛋白含量均比全部底施有不同程度的提高－ 随追氮时期后延,清蛋白含量变化不大,醇溶蛋白含量以抽穗期追氮为最高,球蛋白和麦谷蛋白含量则以孕穗期追氮为最高－ 醇溶蛋白的积累动态呈“高－ 低－ 高”的变化趋势,麦谷蛋白积累则呈现由低到高的变化动态,氮素供应对２ 种蛋白质组分的积累动态有明显的调节效应－     

肾蕨离体培养技术的研究

以肾蕨匍匐茎茎尖为外植体,对离体再生培养技术进行了研究。结果表明：GGB的最佳诱导培养基为MS＋BA0.7 mg.L-1,诱导率为95%;GGB增殖培养基为MS＋BA0.7 mg.L-1,增殖系数为5.3;孢子体诱导培养基为MS＋BA0.7 mg.L-1＋NAA0.7 mg.L-1,诱导率可达到82%;适合诱导生根培养基为MS＋NAA0.2 mg.L-1＋0.1%Ac,生根率为100%。     

Effect of delayed sowing on grain number,grain weight,and protein concentration of wheat grains at specific positions within spikes

Delays in sowing have significant effects on the grain yield, yield components, and grain protein concentrations of winter wheat. However, little is known about how delayed sowing affects these characteristics at different positions in the wheat spikes. In this study, the effects of sowing date were investigated in a winter wheat cultivar, Shannong 30, which was sown in 2019 and 2020 on October 8 (normal sowing) and October 22 (late sowing) under field conditions. Delayed sowing increased the partitioning of ¹³C-assimilates to spikes, particularly to florets at the apical section of a spike and those occupying distal positions on the same spikelet. Consequently, the increase in grain number was the greatest for the apical sections, followed by the basal and central sections. No significant differences were observed between sowing dates in the superior grain number in the basal and central sections, while the number in apical sections was significantly different. The number of inferior grains in each section also increased substantially in response to delayed sowing. The average grain weights in all sections remained unchanged under delayed sowing because there were parallel increases in grain number and ¹³C-assimilate partitioning to grains at specific positions in the spikes. Increases in grain number m^–2 resulted in reduced grain protein concentrations as the limited nitrogen supply was diluted into more grains. Delayed sowing caused the greatest reduction in grain protein concentration in the basal sections, followed by the central and apical sections. No significant differences in the reduction of the grain protein concentration were observed between the inferior and superior grains under delayed sowing. In conclusion, a 2-week delay in sowing improved grain yield through increased grain number per spike, which originated principally from an increased grain number in the apical sections of spikes and in distal positions on the same spikelet. However, grain protein concentrations declined in each section because of the increased grain number and reduced N uptake.     

氮素对稻米蛋白质组分含量及蒸煮食味品质的影响

选用7个粳稻品种,采用裂区试验设计方法研究了氮素对稻米味度值、淀粉谱特性及蛋白质组分含量的影响。结果表明,蛋白质和蛋白质组分含量以及黏滞峰消减值等随氮素营养水平的提高而增加,但大多数供试品种的味度值、下降黏度值和最高黏度反而下降;蛋白质组分百分比组成受氮素营养的影响很小;味度值与蛋白质含量呈极显著负相关,与直链淀粉含量呈不显著负相关;各蛋白质组分含量与稻米味度值的相关均没达到显著水平,除醇溶蛋白含量与最终黏度和回冷黏滞性恢复值呈显著的负相关外,其他蛋白质组分含量与淀粉谱特性中的各项指标间相关均没达到显著水平。     

普通小麦抗赤霉变异细胞系的离体筛选及其再生系抗性分析

通过禾谷镰刀菌毒素胁迫下的幼胚直接筛选、愈伤组织筛选、一步筛选和多步筛选相结合的方法, 从普通小麦千斤早和小偃22中筛选出了抗镰刀菌毒素变异细胞系,比较分析了不同筛选方法在不同筛选程序上的筛选效果,研究了再生株系R0代的抗赤霉特性。结果表明,利用毒素胁迫对优良品种进行抗赤霉细胞工程定向改良是有效可行的,可获得抗病性显著提高的变异株系;利用幼胚直接多步筛选,在高毒素质量浓度上获得抗性植株的比例和抗性提高的幅度均较从愈伤组织筛选和低毒素质量浓度上获得的效果更好。     

体外培养小鼠生精细胞的形态学

利用姬姆萨原液和瑞-姬染色法对30～35日龄小鼠精液和睾丸组织培养后的生精细胞和精子进行形态学鉴定.结果表明,用姬姆萨原液染色鉴定,精子形态清晰,细胞膜边缘清楚,呈紫红色,未成熟精子的染色程度浅于成熟精子;经过培养的睾丸组织用姬姆萨原液染色,生精细胞形态清晰,细胞核呈颗粒状,开始呈现深紫红色,胞质粉红色且带有蓝色背景,颜色较深,一定时间后颜色变浅;瑞-姬染色生精细胞形态完整清晰,细胞核、核仁、胞质分别呈现不同颜色.     

大麦不同粒位粒重和蛋白质含量及发芽整齐度研究

测定了大麦不同穗数株及5个品种（系）不同粒位（顶部、中部和底部）籽粒的粒重、蛋白质含量和发芽率。结果表明：不同穗数株及品种（系）各粒位间籽粒千粒重的差异表现相同的趋势，即中部最大，顶部和底部较小。不同穗数株主茎穗各粒位籽粒蛋白质含量的最大值，1穗株为顶部，2穗株为中部，3穗株则为底部；各品种（系）主茎穗和分蘖穗不同粒位蛋白含量加权平均值均以秀麦3号为最大，97－33最小，且主茎穗各粒位蛋白含量以顶部为最高，不同穗数株各粒位籽粒发芽速度均以顶部最快，中部和底部相对较慢；品系99－18的发芽整齐度最好，吕系92－11发芽最慢且发芽率最低。     

野生花卉胭脂花(Primula maximowiczii)组培体系的建立

以胭脂花(Primula maximowiczii)的种子为外植体,利用单因子和正交试验设计,通过对种子表面消毒、丛生芽诱导和增殖、生根、炼苗和移栽等技术环节的研究,筛选出各培养阶段的最佳培养基配方和培养条件,建立了胭脂花无菌培养体系.结果表明:种子表面消毒的最佳方法是以6％ NaClO消毒6 min;最佳丛生芽诱导增...     

同核异质玉米幼胚的离体培养

本研究选用7对同核异质玉米,进行了幼胚离体培养。研究结果表明:C 细胞质玉米的愈伤组织诱导率明显低于其正常(N)细胞质的保持系。而在同核条件下不同细胞质对愈伤组织的生长速率没有明显影响,但对愈伤组织分化频率影响较大。     

猪体外成熟卵母细胞的电激活及其激活后的体外培养

 主要研究了猪体外成熟卵母细胞电激活的条件以及孤雌激活胚胎的体外培养体系。用不同场强、脉冲时程和脉冲次数激活猪体外成熟的卵母细胞。结果表明 ,在本试验条件下电激活最佳场强和脉冲时程为 130V·mm-1/ 80 μs ,其囊胚发育率为 (2 0 .12± 8.18) % (P >0 .0 5 )。多次脉冲并不比单次脉冲的激活效果好 (P >0 .0 5 )。孤雌激活胚胎用不同培养方式和气体环境在体外培养 7d ,发现 7d不换液与第 5天换液和第 5天换含 10 %胎牛血清的培养液其囊胚发育率 [分别为 (2 5 .30± 7.5 5 ) %、(2 6 .4 4± 8.35 ) %和 (17.6 8± 5 .39) % ]无显著差异 (P >0 .0 5 ) ,后两者换液培养方式其囊胚细胞数 (15 .78± 5 .4 6、14 .5 5± 4 .81)明显低于不换液的 (18.0 1± 6 .79,P <0 .0 1)。同时发现低氧 (5 %CO2 ∶7%O2 ∶88%N2 )气体环境培养的胚胎囊胚发育率和囊胚细胞数和高氧环境 (5 %CO2 的空气 )无明显差别 ,分别为 [(2 0 .78± 8.80 ) %、17.0 0± 6 .12和 (2 5 .30± 7.5 5 ) %、18.0 1± 6 .79,P >0 .0 5 ]。表明激活后第 5天换原液和换含胎牛血清的培养液不能提高囊胚细胞数。低氧 (5 %CO2 ∶7%O2 ∶88%N2 )气体环境培养猪胚胎的效果不比高氧环境 (5 %CO2 的空气 )好。     

Studies on Electrical Activation of Porcine Oocytes Matured in vitro and Embryo Culture Systems

Conditions for electrical parthenogenetic activation of porcine oocytes matured in vitro and in vitro culture systems of porcine embryo were studied. The best results were achieved under the conditions of electrical field strength and the pulse duration at 130Vmm-1/80 μs, with a blastocyst development rate of (20.12 ± 8.18)% (P ＞ 0.05). No significant difference was found between treatments of multiple pulses and a single pulse ( P ＞ 0.05). Parthenogenetic embryos were cultured with different methods and air conditions for 7 days in vitro, blastocyst development rate of embryos with changed culture media [ (26.44 ± 8.35)% ] or changed media with 10% fetal bovine serum (FBS) [ (17.68 ± 5.39)% ] on the fifth day showing no significant difference from that of embryos without change of culture media [ (25.30 ± 7.55) %, P ＞ 0.05 ], while cell numbers of blastocysts from embryos with changed culture media (15.78 ± 5.46 and 14.55 ± 4.81) were significantly lower than number of blastocysts from embryos without change of culture media (18.01 ± 6.79,P ＜ 0.01 ). Blastocyst development rate and blastocyst cell number of embryos cultured in lower O2 (5 % CO2:7%O2:88%N2) also showed no significant difference from those in high O2 (5% CO2 in air) [ (20.78 ± 8.80) % and 17.00 ± 6.12 vs. (25.30 ± 7.55) % and 18.01 ± 6.79, P ＞ 0.05 ]. It is concluded that change of culture media with the same new one or changing over to media with 10% fetal bovine serum (FBS) on the fifth day and low O2 environment are not necessary for porcine embryos development.     

植物离体培养技术生产次生代谢物的研究进展

在植物离体培养中,选择高产的外植体,寻找合适的培养条件,运用不同的离体培养方法是提高细胞生长速度和次生代谢物产量并实现工厂化生产的先决条件.为此,从以上几个方面阐述了近年来的研究进展,着重介绍了8种利用植物离体培养技术生产次生代谢物的方法,并提出了各种方法存在的问题及解决方案,同时对植物离体培养技术的前景进行了展望.