Four freshly established cell lines of blue spruce (Picea pungens Engelm.), with different capacities for somatic embryogenesis (SE), were studied to assess their SE formation and maturation, aiming at identifying key factors responsible for the variation in SE potential. We examined dynamic cytological and physiological changes, including morphology and structure, storage substances, antioxidant enzyme activities and endogenous hormone levels. The results showed that cell lines 202, 201, 211 with SE capacity exhibited active metabolic processes and significant increases in superoxide dismutase (SOD) and peroxidase (POD) activities, total protein (TP) and malondialdehyde (MDA) contents across developmental stages. In contrast, the cell line lacking SE capacity showed low or unchanged enzyme activities. These findings suggest that SOD, POD, MDA and TP can serve as physiological markers for identifying SE capacity. During SE development and maturation, endogenous hormone levels (GA, IAA, ABA) remained unchanged in the non-embryogenic cell line. However, in highly embryogenic line, hormone levels generally increased and then decreased, while lines with intermediate and weak SE showed a steady increase. Our findings provide critical insights into optimizing culture media formulations to enhance blue spruce SE maturation, thereby improving efficiencies of elite genotype selection and large-scale clonal propagation. Such advancements are essential for meeting the growing demand for high-quality planting materials in landscape applications and forestry production. Nevertheless, the underlying molecular mechanisms by which reactive oxygen species (ROS) influence SE maturation capacity, as well as the synergistic regulatory interactions among endogenous hormones during different embryo developmental stages, remain insufficiently understood and warrant further investigation. Furthermore, the technological framework and regulatory patterns identified in this study are not limited to blue spruce; the physiological markers and developmental cues elucidated herein may serve as valuable references for SE research in coniferous species. To validate the practical applicability of this approach, comprehensive field trials are required to evaluate the growth performance, adaptability, and stability of SE-derived plants under natural environmental conditions.

Cranberry(Oxycoccos)has strong cold resistance,and its fruit has high health value and economic value.In this stud-y,the tissue cultured seedlings of cranberry were used as cuttings to explore the soaking and quick dipping tests of cranberry cuttings under different indole-3-acetic acid(IAA),1-naphthlcetic acid(NAA)and 3-indolebutyric acid(IBA)concentrations,and the effects of different hormone types and concentrations on rooting rate,seedling height growth,biomass and root growth characteristics(including the longest root length,root surface area,average root diameter and average root number).The results showed that,when NAA soaking treatment was 100 and 200 mg/L,the root system grew well,but the longest root length was only(30.77±0.18)cm(NAA 100 mg/L),and the average root diameter was(0.3±0.02)mm.However,the longest root length of cuttings treated with IBA 1000 mg/L per hour was(30.13±3.22)cm,the average root diameter was(0.39±0.04)mm,the root surface area was(1.27±0.01)cm2,and the average number of roots was(128.22±2.38).From this point of view,there was little difference in root growth between the soaking treatment and the rapid soaking treatment,but in general,when the IBA concentration was 1000 mg/L,the rooting rate,was 85% at the highest.Therefore,the quick dipping method was more suitable for the rooting of cranberry tissue culture seed-lings.In this study,the cutting propagation technology of tissue culture seedlings of cranberry was established for the first time,which laid the foundation for large-scale propagation of cranberry.

Embryogenic tissue (ET) is important for genetic modification and plant re-generation. The proliferation ability and vigor of ET are crucial for plant propagation via somatic embryogenesis. In this study, ET was induced from mature zygotic embryos in blue spruce (Picea pungens Engelm.). There were significant differences in ET induction between two provenances, i.e. 78.8 ± 12.5% and 62.50 ± 12.8% respectively. Effects of 2,4-Dichlorophenoxy acetic acid (2,4-D), 6-Benzyl amino-purine (6-BA) and/or sucrose on ET proliferation and somatic embryo (SE) maturation were further investigated with four cell lines. The highest ET proliferation rate reached 1473.7 ± 556.0% biweekly. Concentrations of 2,4-D or 6-BA applied at tissue proliferation stage impacted SE maturation among the cell lines, whereas sucrose showed less effects. The highest rate, 408 ± 230 mature SEs/g FW, was achieved in SE maturation cultures. This research demonstrated that the culture conditions, i.e. the specific concentrations of 2,4-D and BA, at ET proliferation stage affected not only ET growth, but also the quality of ET for SE maturation. This study revealed the necessity and benefit in developing both the general and the genotype-specific protocols for efficient production of mature SEs, or somatic plants in blue spruce.

以蔓越莓组培苗为插穗,探究8 种基质对扦插生根的影响.结果表明:全泥炭土不适于组培苗扦插生根,生根率只有22％;总体来看全苔藓基质最适宜组培苗扦插生根,生根率最高,为 86％,苗高增长量为 4.62±0.25 cm,生物量最高(鲜重2.33±0.18 g,干重0.51±0.04 g),生根状况也最好.

Cellular redox state and hormone levels have important regulatory roles in the process of somatic embryogenesis (SE), but experimental data on the topic are limited in conifer SE. This study investigated changes in redox pairs (reduced glutathione (GSH) vs oxidized glutathione (GSSG) and ascorbic acid (ASA) vs dehydroascorbate (DHA)), activities of enzymes (glutathione reductase (GR), DHA reductase (DHAR), and ascorbate peroxidase (APX)), and levels of endogenous hormones (indole-3-acetic acid (IAA), zeatin-riboside (ZR), gibberellin (GA), and abscisic acid (ABA)) following exogenous supplements of 2,4-dichlorophenoxyacetic acid (2,4-D), GSH, and L-buthionine sulfoximine (BSO) to the proliferation medium of Picea pungens SE and linked the changes with proliferation efficiencies. While 2,4-D promoted proliferation and maturation in a concentration-dependent manner, the joint application of 2,4-D (2 mg/L) and GSH (0.5 mmol/L) was the most effective. Such beneficial roles were not observed when GSH alone was applied and exogenous BSO (1 mmol/L) prohibited proliferation. Supplements of 2,4-D and GSH, individually or jointly, enhanced the GR activity, decreased the DHAR activity and increased amounts of redox compounds, yet more in GSH and ASA than in GSSG and DHA, creating reduced environments. Exogenous BSO had reversal effects, creating an oxidized environment, and so was the joint use of 2,4-D and BSO, although at a less extent. 2,4-D and GSH reduced while BSO increased IAA content. The alternations in redox state as well as in IAA content from exogenous 2,4-D or BSO supplements coincided with the corresponding changes in proliferation. The notable finding was that exogenous GSH, regardless its substantial roles in reducing environments or IAA content, did not improve proliferation. Thus, the role of GSH on proliferation varied, depending on 2,4-D existing, and could not be fully accounted for by dynamic changes in redox state or hormone level. Alternations in APX and hormones ABA, ZR, GA, and IAA/ABA ratio were also found not highly involved in altering proliferation.

以吉林省西部轻度盐碱地种植的沙枣、枸杞、蒙桑和柠条锦鸡儿实生苗为试验材料,针对4种植物的光合参数和叶绿素荧光参数等相关方面进行差异性和相关性分析,揭示4种植物的光合性能,以期为盐碱地治理提供参考.结果表明,4种植物的光合特征和叶绿素荧光特性差异显著,净光合速率(Pn)由大到小依次为:枸杞、蒙桑、沙枣、柠条.试验植物叶片的净光合速率(Pn)与气孔导度(Gs)有显著的正相关性,因而Pn受Gs影响较大;并且Pn与光适应下的最大光化学效率、实际光化学效率(ФPSⅡ)、光化学淬灭系数(qP)和电子传递效率(ETR)等荧光参数密切相关.枸杞的J相和I相的相对可变荧光速率(Vj和Vi)最低,质体醌电子传递效率高,光系统(PSⅡ)受体侧的质体醌库(Sm)最大,光反应活性最大.柠条电子传递效率低,光反应活性受到天线色素对能量(ABS)的吸收和反应中心能量捕获(TR)的限制.同时,枸杞的性能指数(PI ABS)在以初始荧光(Fo)为基础的性能指数(PICSo)和以最大荧光(Fm)为基础的性能指数(PICSm)显著高于其他树种,柠条最低.4种植物的3个性能指数之间具有显著差异性和极显著相关性,且Pn与PI ABS、PI CSo、PI CSm呈显著相关性.因此,PI ABS、PI CSo和PI CSm适合作为4种植物叶绿素荧光特性的评价指标.研究表明,4种植物中枸杞的性能指标(PI)和Pn最高,因而光合性能最强,蒙桑和沙枣次之,柠条最低.所以枸杞的光合生理活性强,适合本试验盐碱地环境的栽植和引种.

Picea pungens (Engelm.), known for its blue-green needles, has become a likable ornamental species in northeast China since 2000. Nonetheless, a lack of propagation methods that can maintain genetic fidelity and develop seedlings at a large scale prevents the further expansion of the species. Somatic embryogenesis (SE), paired with cryopreservation technologies, may provide a valid alternative. Picea pungens SE is not new, but its practical application has been limited due to low efficiencies in SE initiation and maturation as well as a lack of effective cryopreservation technology. In this study, experiments were carried out to overcome the limitations by modifying culture media. For initiation, the efficiency was enhanced by adjusting concentrations of 2.4-dichlorophenoxy acetic acid (2,4-D), 6-benzyl amino–purine (6-BA) or sucrose supplemented to the induction medium. The concentrations of 4.0 mg/L 2,4-D, 2 mg/L 6-BA, and 5 to 10 g/L sucrose were found optimal in maximizing initiation efficiency. For maturation, the efficiency, expressed as the number of mature somatic embryos per gram of fresh mass cultured (E/gFM), varied greatly with the choices of the basal medium and concentration of abscisic acid (ABA) of the maturation medium. Based on our results, the judicial choices were using the DCR medium as the basal medium and 10 mg/L ABA. The maturation efficiency could also be improved by adjusting the maturation medium’s osmotic pressure by manipulating the concentrations of carbohydrate and Gelrite and culture density. While the maturation medium, using sucrose as carbohydrate source or supplemented with a low (<8 g/L) Gelrite concentration, facilitated maturation, optimal selections were truly genotype-dependent. Our results also suggest that, while the optimal culture density varied with genotype, in general it is needless to culture more than 100 mg embryogenesis tissues per dish (size: 10 × 1.5 cm). Based on this study, the optimum pretreatment for embryogenesis tissue cryopreservation was culturing the tissues on the proliferation medium with 0.4 mol/L sorbitol for 24 h, followed by treatment with 5% Dimethyl sulfoxide. This study significantly improved the initiation (achieved a frequency of 0.56) and embryo maturation efficiencies (achieved 1030 E/gFM) and established an effective preculturing protocol for cryopreservation (recovered 1354 E/gFM) for the species. The protocols developed here, paired with the available ones for other SE steps in the literature, form a well-refined SE technology intended for commercial application to Picea pungens.

以耐寒月季'高山朝霞'嫩枝为材料,研究不同扦插基质、不同生长调节剂种类(IBA、NAA、IAA)和质量浓度(400 mg·L-1、800 mg·L-1、1200 mg·L-1)对其扦插生根的影响.结果表明:最适合的基质为河沙+蛭石+珍珠岩(体积比3:1:1),生根率达到93.33％.经不同生长调节剂处理后,生根率较对照有显著提高,1200 mg·L-1 IBA处理后的根系生长和根系构型最佳.

[目的]通过对红皮云杉(Picea koraiensis)体细胞胚胎发生条件的筛选和超低温保存技术研究,建立其完整的体胚发生体系及超低温保存条件,为红皮云杉优良种质资源的大量繁殖及保存提供基础.[方法]以红皮云杉合子胚为外植体,以两种基本培养基、6种植物生长调节剂组合为培养条件,筛选胚性愈伤组织诱导的适宜条件.并将获得的胚性愈伤组织进行超低温保存和体胚发育与成熟萌发试验,以3种脱落酸(ABA)和Gelrite浓度以及4种基本培养基为培养条件,筛选出体胚成熟和萌发的适宜培养条件.[结果]①两种基本培养基相比较,改良RJW(诱导率为30.00％)优于1/2 LV(诱导率为22.50％)基本培养基.当萘乙酸(NAA)质量浓度为3.0 mg/L、6-苄氨基腺嘌呤(6-BA)质量浓度为0.5 mg/L时胚性愈伤组织诱导率最高为50.00％;②胚性愈伤组织增殖培养3个月后进行超低温保存试验,经恢复培养10 d左右可以清晰地观察到成活的胚性愈伤组织,3个细胞系的胚性愈伤组织均可成活;③体胚发育阶段,改良RJW培养基添加Gelrite 6.0 g/L和ABA 20 mg/L时,3个细胞系体胚数量均较高,但是畸形胚比例也偏高,特别是HY-1细胞系畸形胚比例高达77.14％;当Gelrite质量浓度增加到8.0 g/L时,3个细胞系畸形胚比例均降低,其中HY-2细胞系体胚数量为396.00个/g,畸形胚比例为10.33％;④红皮云杉体胚成熟培养两个月进行体胚萌发,在1/2 LM上体胚萌发能力最弱,其次是LM基本培养基,改良RJW体胚萌发能力最佳.不同细胞系体胚萌发能力也不同,体胚萌发能力最高为65.00％.[结论]① 胚性愈伤组织诱导的适宜培养基为改良RJW基本培养基,适宜的激素组合为NAA 3.0 mg/L+6-BA 0.5 mg/L;②本试验进行超低温保存的细胞系均可成活;③体胚发育与成熟适宜的ABA质量浓度为20 mg/L,适宜Gelrite质量浓度为8.0 g/L;④体胚萌发的适宜基本培养基为改良RJW基本培养基.本研究建立了红皮云杉体胚发生体系并成功进行胚性愈伤组织的超低温保存条件,该技术体系可用于红皮云杉优良种质的快速繁殖.

[目的]为探讨盐碱胁迫条件下沙枣幼苗的生长与光合特性,以掌握沙枣苗期不同胁迫条件下的抗逆性,为培育沙枣耐盐碱品种和改进栽培管理措施提供理论基础.[方法]选取1年生长势较一致的沙枣幼苗为试验材料,采用盆栽试验并进行随机区组设计.试验共分4个试验组和1个对照组,以200 mmol·L-1、pH 8.07～10.29的碱性复合盐为处理,对照用清水处理.6个试验周期后对其生长指标与光合特性进行了初步测定.[结果]相同Na+浓度的碱性复合盐胁迫条件下,沙枣幼苗的光合参数出现明显变化,其净光合速率(A)、蒸腾速率(E)、气孔导度(gsw)分别随胁迫处理pH值升高而下降,苗高生长量、地径生长量、新梢生长量下降明显;在Ph值达到8.67以后,随着NaHCO3、Na2CO3比例增加,生长显著下降.相关分析表明3个生长指标与净光合速率(A)呈极显著正相关(P<0.01),并且均与蒸腾速率、气孔导度呈显著正相关(P<0.05).通过光和特性与生长指标的隶属函数综合分析表明,随着pH值的升高,碱性复合盐对沙枣苗期的胁迫程度增强.[结论]在相同Na+浓度碱性复合盐胁迫条件下,盐碱胁迫效应显著.在本试验范围内,200 mmol·L-1的碱性复合盐不同处理条件下,随着处理组pH的增大,生长与光合指标呈下降趋势;当pH值升至8.67以后,各生长指标下降明显,从而说明了高pH碱性复合盐胁迫条件下,沙枣幼苗对盐的耐受程度强于碱,但碱性盐胁迫更强.

Abstract This study developed somatic embryogenesis protocols for Picea pungens (Engelm), an important ornamental species, including initiation, proliferation, maturation, germination, and acclimation. Somatic embryogenic tissues were induced from mature zygotic embryos of five families, with a frequency of $$\ge $$ ≥ 22% for each. Embryogenic tissues (ET) from 13 clones of three families were proliferated for one week, achieving an average rate of 179.1%. The ET of 38 clones of three families were cultured in maturation medium for six weeks; 188 mature embryos on average were counted per gram ET cultured, of which $$\ge $$ ≥ 81.1% appeared normal, and each clone developed at least 28 normally matured embryos. A total of 69.9% or more of cotyledonary somatic embryos germinated normally and developed into normal emblings. The experiment of transplanting the emblings into a greenhouse had an average survival rate of 68.5%. Considerable variation among and within families during initiation and proliferation was observed, but this variation decreased in the maturation and germination. Changing the concentration of plant growth regulator of the initiation medium did not significantly change the initiation frequency. We recommend incorporating these protocols into the current Picea pungens practical programs, although further research is essential to increase efficiencies and reduce cost.

红松是重要的用材树种和坚果经济林树种.以组织培养的方法,选用三岔子红松种子园5个家系的未成熟种子作为外植体进行胚性愈伤组织诱导研究.结果 表明,不同家系未成熟种子在添加萘乙酸(NAA)2 mg/L+6-苄氨基嘌呤(6-BA)1 mg/L的培养基上胚性愈伤组织诱导率不同,最佳诱导率为5.68％.红松愈伤组织诱导的最适蔗糖浓度为30 g/L.选取长势良好的胚性愈伤组织进行超低温保存,在恢复培养10 d后可观察到不同细胞系均有明显新生愈伤组织,15 d后愈伤组织大量增殖.本研究为三岔子红松种子园红松优良种质资源的规模化繁殖和保存奠定了基础.

本文对从俄罗斯引进的24个花楸品种,采取嫁接和组织培养方式进行保存.初步分析各品种的保存和生长情况,认定花楸品种可采用嫁接和组织培养方法进行保存和繁殖,为今后花楸品种的筛选和繁殖提供方法.

以唐古特白刺1年实生苗为研究对象,采用盆栽试验,以浓度200 mmol·L-1、pH 8.07～10.29的碱性复合盐NaCl、Na2 SO4、NaHCO3、Na2 CO3为处理,探究盐碱胁迫对唐古特白刺幼苗光合特性及叶片部分生理特性的影响.结果表明:相同Na+浓度不同盐分配比胁迫条件下,唐古特白刺幼苗叶片丙二醛(MDA)含量、胞间CO2浓度(Ci)随胁迫处理pH升高而增加,而叶绿素含量(Chl)、含水率(LWC)、净光合速率(A)、蒸腾速率(E)、气孔导度(gsw)分别随胁迫处理pH升高而下降;在pH达到8.67以后,随着NaHCO3、Na2 CO3摩尔分数增加,幼苗叶片Chl、A显著下降,充分说明,相同Na+浓度下,盐碱胁迫效应显著,但碱性盐胁迫更强.

以红皮云杉作为砧木,北美蓝云杉当年生枝条为接穗,探究不同嫁接方法和时间对嫁接效果的影响.结果表明:5月中旬以切接的嫁接方法更适合于北美蓝云杉异砧嫁接,嫁接成活率达81％.

[目的]探讨黑果枸杞的组织培养技术,为实现规模化高效离体快繁提供技术依据.[方法]以黑果枸杞当年生嫩茎为外植体,研究激素组合对黑果枸杞组培苗初代、增殖以及生根培养的影响.[结果]适合黑果枸杞初代培养的培养基为MS+6-BA1.0 mg/L+NAA0.2 mg/L,腋芽生长良好,萌芽率达88.61％,且玻璃化情况较少;适合微枝增殖培养的培养基为MS+6-BA0.1mg/L+NAA0.2mg/L,微枝生长速度快,增殖倍数可达4.8倍,少见玻璃化现象,且苗生长健壮,叶片嫩绿;适合微枝生根的培养基为1/2MS+IBA0.2mg/·L,生根率达98％.适宜的移栽基质为营养土:河沙:蛭石=3:1:1(体积比).[结论]黑果枸杞嫩茎离体培养和茎芽增殖可以获得再生植株实现离体快繁,本研究结果为黑果枸杞优良种质规模化繁殖提供了技术支持.

以欧洲花楸成熟种子为外植体,研究激素组合对欧洲花楸培养基内萌发、增殖以及生根培养的影响,建立组培快繁体系.结果表明:适合欧洲花楸种子萌发阶段培养的培养基为MS+6-BA0.5 mg·L-1+NAA0.1 mg·L-1,萌芽率达68.76％;适合组培苗增殖培养的培养基为WPM+6-BA0.4 mg·L-1+NAA0.05 mg·L-1,增殖倍数可达7.70倍;生根培养最适培养基为1/2MS+NAA0.3 mg·L-1+IBA0.1 mg·L-1,平均生根率达65.21％.

体胚发生具有遗传稳定、繁殖快、数量多结构完整等优点,为云杉属树种细胞分化和全能性表达提供较为理想的理论体系.体胚发生的影响因素主要有,外植体的部位及所处生理时期、基本培养基的种类、植物生长调节物质的种类与浓度、糖的种类与浓度、以及培养条件等,不同因素对体胚发生的影响也不尽相同.本文综述了云杉属树种体胚发生的研究进展.

In order to explore the influence factors of the proliferation and the mature stage of somatic embryo of Picea crassifolia, the mature embryo was used as explants, and the related research of somatic embryos was carried out. The results showed that: the best medium for the callus maintenance and proliferation stage of the Picea crassifolia was 1/2 LV + BA 1.1 mg/L+ 2,4-D 3.3 mg/L + 10g/Lsucrose + L-glutamine 400 mg/L + hydrolyzed casein 400 mg/L + Phytagel 3.5g/L. The best subculture cycle of embryogenic callus was about 14-16 days. During the development and maturation of somatic embryos, when the ABA concentration was 30 mg/L, the number of normal embryos obtained by two seed germinal callus was the most.