Pectin methylesterase inhibitors GhPMEI53 and AtPMEI19 improve seed germination by modulating cell wall plasticity in cotton and Arabidopsis

The germination process of seeds is influenced by the interplay between two opposing factors,pectin methylesterase(PME)and pectin methylesterase inhibitor(PMEI),which collectively regulate patterns of pectin methylesterification.Despite the recognized importance of pectin methylesterification in seed germination,the specific mechanisms that govern this process remain unclear.In this study,we demonstrated that the overexpression of GhPMEI53is associated with a decrease in PME activity and an increase in pectin methylesterification.This leads to seed cell wall softening,which positively regulates cotton seed germination.AtPMEI19,the homologue in Arabidopsis thaliana,plays a similar role in seed germination to GhPMEI53,indicating a conserved function and mechanism of PMEI in seed germination regulation.Further studies revealed that GhPMEI53 and AtPMEI19 directly contribute to promoting radicle protrusion and seed germination by inducing cell wall softening and reducing mechanical strength.Additionally,the pathways of abscicic acid(ABA)and gibberellin(GA)in the transgenic materials showed significant changes,suggesting that GhPMEI53/AtPMEI19-mediated pectin methylesterification serves as a regulatory signal for the related phytohormones involved in seed germination.In summary,GhPMEI53 and its homologs alter the mechanical properties of cell walls,which influence the mechanical resistance of the endosperm or testa.Moreover,they impact cellular phytohormone pathways(e.g.,ABA and GA)to regulate seed germination.These findings enhance our understanding of pectin methylesterification in cellular morphological dynamics and signaling transduction,and contribute to a more comprehensive understanding of the PME/PMEI gene superfamily in plants.

Arginine promotes seed energy metabolism,increasing wheat seed germination at low temperature

Low temperatures during germination inhibit seed growth,lead to small and weak seedlings,and significantly reduce the wheat yield.Alleviating the adverse effects of low temperature on wheat seed germination is highly important for achieving high and stable wheat yields.In this study,Tongmai 6(insensitive)and Zhengmai 113(sensitive),which have different low-temperature sensitivities during germination were treated with low temperature during germination.The transcriptome,metabolome and physiological data revealed that low temperature decreased the germination rate,downregulated the expression of a large number of genes involved in regulating glycometabolism,and inhibited carbon,nitrogen(especially amino acids)and energy metabolism in the seeds.Arginine content increased at low temperature,and its increase in the low-temperature-tolerant variety was significantly greater than that in the sensitive variety.Arginine priming experiment showed that treatment with an appropriate concentration of arginine improved the seed germination rate.The conversion of starch to soluble sugar significantly increased under exogenous arginine conditions,the content of key metabolites in energy metabolism increased,and the utilization of ATP in the seeds increased.Taken together,arginine priming increased seed germination at low temperature by relieving inhibition of seed carbon and nitrogen metabolism and improving seed energy metabolism.

Genome-wide association study reveals that JASMONATE ZIM-DOMAIN 5 regulates seed germination in rice

Seed germination is a complex trait regulated by multiple genes in rice.However,the regulators of rice seed germination have yet to be sufficiently determined.Here,a quantitative trait locus(QTL)for rice seed germination was identified in a genome-wide association study.The candidate gene JASMONATE ZIM-DOMAIN 5(OsJAZ5)of the QTL was verified that positively regulates seed germination.OsJAZ5 regulation of seed germination involves an OsABI3-mediated abscisic acid pathway.Overexpression of OsJAZ5 facilitated seed germination.The application of OsJAZ5 might be useful for increasing seed germination for rice direct seeding.

Preparation of Tartary Buckwheat Seed Coating Agent and Its Effect on Germination

To mitigate the wastage of seed resources and reduce the usage of pesticides and fertilizers, seed coating agentshave gained popularity. This study employs single-factor and multi-index orthogonal experimental design methodsto investigate the seed coating formula and physical properties of Tartary buckwheat. The specific effects ofeach component on Tartary buckwheat seed germination are analyzed. The findings reveal that the seed coatingagent formulated with 1.5% polyvinyl alcohol, 0.15% sodium alginate, 0.2% op-10, 0.1% polyacrylamide, 8% colorant,3% ammonium sulfate, 1% potassium dihydrogen phosphate, and 0.15% carbendazim exhibits the mosteffective coating. It demonstrates optimal physical properties and promotes seed germination efficiently. The suspensionrate of this seed coating agent reaches 91.12%, with a mere 2.13% coating shedding rate and 2.5% coatingseed rot rate. Furthermore, it achieves a germination percentage of 99.17%, which is 20.84% higher than the lowestgroup. The germination potential and index are also significantly higher than the lowest group, with anincrease of 20.84% and 26.56%, respectively. Additionally, the vitality index is 553.08, a 15.75% increase comparedto the lowest group. The application of seed coating agents helps reduce seed resource loss, increase plant numbers,and ultimately enhance agricultural yields. This finding holds practical significance in agriculturalproduction.

Effects of Low Temperature Stress on Germination and Physiological Characteristics of Different Sweet Maize Varieties

[Objectives]The paper was to explore the effects of low temperature stress on germination and physiological characteristics of different sweet maize varieties.[Methods]Taking Taitian 264,Zhexuetian 1 and Chaotian 4 as the research objects,the changes in germination potential,germination index,plant height,biomass,and antioxidant enzyme activity of maize seeds were studied under optimal temperature conditions(25℃)and low temperature stress conditions(10℃).[Results]Under 10℃stress,the germination rate and germination index of Taitian 264 were higher than that of Zhexuetian 1 and Chaotian 4.Under low temperature stress,Taitian 264 exhibited the least reduction in height and biomass,while Zhexuetian 1 had the most reduction.Additionally,the SOD and POD activities of Taitian 264 were higher than that of Zhexuetian 1 and Chaotian 4 under both temperature conditions,while the MDA content of Taitian 264 was lower.Taitian 264 showed strong germination ability against low temperature stress.[Conclusions]This study provides a basis for timely sowing practices of sweet maize in agricultural production.

Effects of Different Exogenous Substances on Seed Germination of Isatis indigotica Under Drought Stress and Chemical Composition of Isatis indigotica leaves

This study was to investigate the effects of three exogenous substances on chemical constituents of Isatis indigotica leavesand their efficacy in alleviating drought stress, and explore the methods of applying exogenous substances to efficient cultivationof Isatis indigotica. Polyethylene glycol (PEG) was used to simulate drought stress to deal with seeds of Isatis indigotica at thegermination stage (concentration: 0, 10%, 15%, and 20%). Simultaneous operation of exogenous growth regulators [microbialinoculum (MI), γ-aminobutyric acid (GABA) and salicylic acid (SA)] and PEG were implemented in seeds of Isatis indigotica.The effects of drought stress and the mitigation of exogenous substances were observed by statistics of seed germination potential,germination rate, hypocotyl length, and radicle length of each treatment. The effects of exogenous substances on the content ofalkaloids, crude protein and free amino acids in the leaves of Isatis indigotica grown in a greenhouse were determined after sprayingexogenous substances on the plants. The differences of germination potential, germination rate, hypocotyl length, and radicle lengthamong 15% PEG stress treatment, 10% PEG stress treatment and the control were significant (P<0.05). According to the predesignedgermination standard, the seeds did not germinate under 20% PEG stress treatment. When the PEG concentration was 15%, the resultsof seed germination potential and germination rate after adding MI were significantly different from those under stress alone (P<0.05).When exposed to 10% PEG stress, the supplementation of GABA led to a notable increase in radicle length of Isatis indigotica seeds,showing significant differences compared to other three treatments. The application of MI and GABA under 15% PEG stress resultedin a significant increase in the radicle and hypocotyl length of Isatis indigotica seeds compared to other two treatments. The contentof the total alkaloids in leaves of Isatis indigotica was significantly increased after spraying GABA. Meanwhile, the contents of crudeprotein and the total free amino acids were kept constant after spraying exogenous substances. Application of MI and GABA couldalleviate drought stress of Isatis indigotica. The content of the total alkaloids in leaves of Isatis indigotica could significantly increaseafter spraying GABA.

Impact of ZnSO_(4)^(-)and ZnO Nanoparticles on Seed Germination and Seedling Growth of Lettuce

Micronutrient nanoparticles(NPs)are currently an option for chemical fertilization and biostimulation in crops.However,there is little information on the phytotoxic or biostimulatory effects of NPs at low concentrations of some elements,such as Zn.In this study,the effect of low concentrations of Zn oxide(ZnO)NPs on germination,growth variables,and nutritional attributes of lettuce(Lactuca sativa L.)was evaluated in comparison to Zn sulfate.Romaine lettuce seeds were treated with ZnSO_(4)^(-)×7H_(2)O and ZnO NPs at Zn molar concentrations of 1×10^(−3),5×10^(−3),1×10^(−4),5×10^(−4),1×10^(−5),5×10^(−5),1×10^(−6),and 5×10^(−6).The seeds treated with ZnSO4−at 5×10^(−6)registered the highest radicle length,73%more than the control treatment.The seeds treated with ZnSO4−at 5×10^(−3)registered the lowest values,with 50%less than the control treatment.ZnO NPs at 5×10^(−6)significantly increased content of chlorophyll A and B and total phenolics.These results indicate the possible existence of a mechanism related to the intrinsic nanoparticle properties,especially at low concentrations.

Influence of Plant Growth Regulators on Pollen Germination and Growth of Xinjiang Apricots

[Objective] This study aimed to investigate the influence of different plant growth regulators on apricot pollen germination and pollen tube growth. [Method] Pollens of six kinds of Xinjiang apricots were cultured in solid media supplemented with five plant growth regulators （GA3 , NAA, 2, 4-D, 6-BA, IAA）. Then the rate of pollen germination and the length of pollen tube were respectively measured. [Result] In a certain concentration range, GA3 most significantly promoted the pollen germination and the pollen tube growth of Shushanggan, Kalayulvke, Dayoujia, Yiliakeyulvke and Kabakehuanna; NNA had the strongest improvement function on Kumaiti’s pollen germination and pollen tube growth. [Conclusion] All the five plant growth regulators promoted the pollen germination and the pollen tube growth of apricots at low concentration but inhibited them at high concentration.

Effects of Pb Stress on Seed Germination and Seedling Growth of Two Weed Species Seeds

Objective] The aim was to investigate the effects of Pb stress on seed germination and seedling growth of Chenopodium glaucum and Chenopodium serot-inum L. [Method] With C. glaucum and C. serotinum as the study objects, the ef-fects of different concentrations of Pb on their seed germination and seedling growth, as wel as on the leaf SOD, POD and CAT activity were studied primarily. [Result] The germination potential of both the two kinds of weed seeds increased first and then decreased with the increase of Pb concentration. The germination po-tential of C. serotinum and C. glaucum seeds reached the peak at Pb concentration of 25 and 50 mg/L, respectively. With the increase of Pb treatment concentration, the germination rate, germination index and vigor index of C. serotinum and C. glaucum seeds decreased, and their bud length and root length also reduced gradu-al y. ln overal , the leaf SOD, POD and CAT activity of C. glaucum and C. serot-inum_ seedlings increased first and then decreased with the increased treatment concentration of Pb. The leaf SOD and POD activity of C. glaucum and C. serot-inum seedlings reached the maximum at Pb concentration of 200 and 100 mg/L, respectively, and the leaf CAT activity reached the maximum at Pb concentration of 100 mg/L. [Conclusion] The two Chenopodium species had strong tolerance to Pb stress, and they could be used as alternative accumulator plants of soil Pb contami-nation.

Effects of Cadmium Stress on Seed Germination and Seedling Growth of Brassica napus L.

To investigate the effects of the heavy metal cadmium （Cd） on seed ger- mination and seedling growth of Brassica napus L., 25 B. napus lines with different genetic background were treated with four concentrations （0, 1, 5 and 10 mg/L） of Cd, and then, their germination potential, germination rate, seedling length, main root length, seedling fresh weight, and seedling dry weight were measured. The re- sults showed that the response of the B. napus lines to Cd stress was different. With the increase of Cd concentration, the relative germination potential and relative seedling rate increased first and then decreased, and reached the maximum levels when the Cd concentration was 5 rag/L. The inhibitory effects of Cd stress on seedling growth indices in a decreasing order were root length〉seedling length〉 seedling fresh weight〉seedling water content. There were extremely significant differ- ences in the relative root length, relative seedling length, relative seedling fresh weight and relative seedling water content among the B. napus lines treated by dif- ferent concentrations of Cd, while there were no significant differences in the rela- tive germination potential and relative seedling rate among the lines, except that the relative seedling rate was significantly different among the lines treated by 5 mg/L Cd. The results suggested that seedling growth was more sensitive than seed ger- mination to heavy metal stress.

Effects of Exogenous GA_3 on Seed Germination and Seedling Growth of Salicornia europaea L. under Salt Stress

[Objective] The paper was to study the effects of exogenous GA3 on seed germination and seedling growth of Salicornia europaea L.under salt stress.[Method] With the combination of different concentrations of NaCl（50,100,150,200,250,300,350,400 mmol/L）with 10 μmol /L GA3 treatments,the effects of exogenous GA3 on seed germination and seedling growth of S.europaea under salt stress were studied.[Result] Exogenous GA3 promoted the seed germination rate,germination capacity,germination index and seed vigor index of S.europaea under salt stress.Under the treatment of exogenous GA3,the root system vigor,the lengths of roots and stems and the salt-tolerance of S.europaea were higher than that of single salt stress treatment.[Conclusion] Within a certain concentration range,exogenous GA3 could alleviate the inhibition effects of salt stress on seed germination and seedling growth of S.europaea.

Effects of Liquid Seaweed Bio-organic Fertilizer on Seed Germination and Seedling Growth of Vegetables

[Objective] This study aimed to investigate the effects of liquid seaweed bio-organic fertilizer on seed germination and seedling growth of different vegetables. [Method] Serial dilution concentrations of liquid seaweed bio-organic fertilizer were prepared for seed soaking and pot incubation of cucumber, tomato and chili, to ob- serve the effects of liquid seaweed bio-organic fertilizer on seed germination and seedling growth of vegetables. [Result] Compared with the control, germination rate of cucumber and tomato seeds applied with 600-fold liquid seaweed bio-organic fer- tilizer varied significantly; germination rate of chili seeds applied with 400-fold liquid seaweed bio-organic fertilizer varied significantly; germination energy and germination index of chili seeds applied with different dilution concentrations of liquid seaweed bio-organic fertilizer presented no significant differences. In addition, 200-fold and 400-fold liquid seaweed bio-organic fertilizer significantly improved the root length, plant height, plant fresh weight, plant dry weight, chlorophyll content and leaf area of cucumber, tomato and chili seedlings; after treated with 600-fold liquid seaweed bio-organic fertilizer, root length, chlorophyll content and leaf area of cucumber seedlings varied significantly compared with the control, but no significant differences were observed in plant height, plant fresh weight and plant dry weight; after treated with 600-fold liquid seaweed bio-organic fertilizer, root length, chlorophyll content, plant height, plant fresh weight and plant dry weight of chili and tomato varied sig- nificantly compared with the control, but no significant differences were observed in leaf area. [Conclusion] Soaking vegetable seeds with liquid seaweed bio-organic fer- tilizer can significantly improve seed generation rate and seedling growth.

Effects of Salt Stress on Seed Germination and Seedling Growth in Raphanus sativus L.

[Objective] This study aimed to investigate the effects of salt stress on seed germination and seedling growth in radish（Raphanus sativus L.）.[Method] First,effects of soaking seeds with clear water and different concentrations of saline solution were compared; then,different concentrations of salt solution （0.2％-1.0％） was used to treat radish during the whole germination period.[Result] soaking seeds with clear water can improve the germination indicators of radish seeds exposed to high salt stress; low salt concentration（0.2％,0.4％） had little effect on the growth indicators of radish seedlings,and 0.2％ salt concentration can even promote the growth of radish seedlings,however,the high salinity had a bad inhibition on seedling growth,the higher the salt concentration,the stronger the inhibition.[Conclusion] According to the comprehensive comparison,the order of the six varieties in salt-tolerance was as follows：Xinxiayuanbai＞W1029,Xiayuanbai ＞ Nanyangbailizhi,Chunyidianhong＞ Sijihong 3.

Inhibition of Chlamydospore Germination and Mycelial Growth of Trichoderma spp. by Chemical Fungicides

For exploring the influences of application and residue of chemical fungi- cides on chlamydospore preparations of Trichoderma spp., the effects of seven chemical fungicides on chlamydospore germination and mycelia growth of two bio- control fungi T. harzianum 610 and T. Iongibrachiatum 758 were studied. Carben- dazim, tebuconazole and difenoconazole showed strong toxicities, thiram and car- bexin showed moderate toxicities, and metalaxyl showed Mycelia growth of the two strains was more sensitive to most tested fungicides than those of chlamydospore germination. Chlamydospore germination of 610 was more sensitive to tested fungicides than those of 758, and mycelia growth of 758 was more sensitive to most tested fungi- cides than those of 610. Among the seven fungicides, 98% carbendazim had the strongest effects （ECru values were 1.64 and 0.05μpg/ml）, and 70% pentachloroni- trobenzene had the weakest effects （EC50 values were 1.64 and 0,05 μg/ml） for chlamydospore germination and mycelia growth of 610. As for 758, 98% carbendaz- im had the strongest inhibitory effects and 95% metalaxyl had the weakest inhibitory for chlamydospore germination of 756 （EC50 values were 0.62 and 1 108.61 μg/ml respectively）, whereas 96.2% tebuconazole showed the strongest inhibitory effects for mycelia growth of 758 （EC= value was 0.32μg/ml）, and 95% metalaxyl was the weakest （EC= value was 1 206.29 μg/ml）. According to the applied concentration of different fungicides in practice, we concluded that chlamydospore preparations of 610 and 758 could not be combined with carbendazim, tebuconazole, thiram and carboxin for controlling plant diseases, and the pesticide residues to the biocontrol effects should be kept in mind. Chlamydospore preparations of 610 and 758 can be and difenoconazole for controlling plant dis- eases, 758 chlamydospore preparations and germinated chlamydospore of 610 can be combined with metalaxyl for controlling plant diseases, and pesticide residue risk was not serious.

Effects of Various Phytohormones on Seed Germination and Seedling Growth of Leymus Chinensis

[Objective] To explore the effects of various phytohormones at different concentrations on the seed germination and seedling growth of Leymus Chinensiss （ L. chinensis）. [ Method ] The seeds of two varieties of L. chinensis were respectively treated with four kinds of phytohormone [ gibbereUin （ GA ）, 2,4-D, naphthalene acetic acid and 6-benzyl aminopurine （6-BA） ] for the observation of seed germination. The seedling growth of natural L. chinensis was observed after the seedlings were respectively treated with these phytohormones. [Result] The germination percentage of L. chinensis was promoted by the GAs and 6-BA at low concentration and inhibited by these two kinds of phytohormone at high concentration. Moreover, they promoted the seedling growth of L. chinensis, and the GA3 had better effect. However, the NAA and 2,4-D at any concentration inhibited the seed germination and seedling growth of L. chinensis. [ Conclusion] The various phytohormones have different effects on the seed germination and seedling growth of L. chinensis.

Study on Cadmium Tolerance of Seed Germination and Seedlings Growth of Different Rice Varieties

The research used bioassay method to explore seed tolerance of Cd in terms of seed germination and seedling growth. Specifically, 12 rice varieties of three types were investigated under Cd stress, covering germination rate, shooting index, vigor index, seedling and shoot lengths, and fresh weights. The results showed that as Cd concentration grew, seed germination and seedling growth kept increasingly inhibited; germination index, vigor index, root length, shoot length and fresh weights went down; inhibition effects on seed germination and seedling growth were conventional rice〉two-line hybrid rice〉three-line hybrid ,rice. Shanyou 9588, Tianyou 998, Chuanxiang 8, and Y Liangyou 1 performed ~rongly in Cd tolerance; C Liangyou 651, Nei 5 You 263, Xiangwanxian 16, Huanghuazhan, Xiangwanxian 13, and Zhunliangyou 527 performed poorly in Cd tolerance, and Cd tolerance of rest treatments were moderate.

Effects of Bamboo Vinegar on Seed Germination and Seedling Growth of Different Flue-cured Tobacco Varieties

[Objective] The present experiment was conducted to study the effects of bamboo vinegar on seed germination and seedling growth of different flue-cured to- bacco varieties, so as to provide some references for its application in tobacco pro- duction. [Method] By using indoor culture dish method and greenhouse seedling- raising plate culture,four different concentrations of bamboo vinegar dilution （50, 100, 200 and 400 times）were designed and clean water was used as the control （CK）. The germination indices of Yunyan 97, Guiyan 2, Jiucaiping 2 and Bina 1 were de- termined after seed germinating. The seedling growth indices of Yunyan 97 were investigated after 10 days of spaying bamboo vinegar on seedling leaves at the 3r~ time. [Result] Compared with CK, different bamboo vinegar dilutions could inhibit the seed germination and seedling growth of all flue-cured tobacco varieties. The seed germination rate and index, vigor index, whole length, root length, stem length and fresh weight of seedlings of all varieties presented decreasing trend to varying de- grees, moreover, most of them reached significant level. Compared with CK, spray- ing different concentrations of bamboo vinegar dilution at the seedling stage could improve the quality of seedlings. In all bamboo vinegar treatments, the plant height, the maximum leaf length and root volume under 100 times dilution treatment were the highest with 3.36 and 10.14 cm, and 0.92 ml, respectively. And the stem girth, dry weight and the maximum leaf width of seedlings under 400 times dilution treat- ment also showed the highest with 1.82 cm, 0.262 g and 5.74 cm, respectively. [Conclusion] Supplying different concentrations of bamboo vinegar solution on culture dish could inhibit the seed germination and seedling growth of flue-cured tobacco, while spraying 100-400 times bamboo vinegar solution to leaves could promote the seedling growth and improve the quality of Yunyan 97 to varying degrees.

Effect of Different Treatments on the Germination and Seedling Growth of Firmiana simplex

With Firmiana simplex （L.） W. F. Wight as an experiment material, the effects of different concentrations of GA3 （Gibberellin A3）, SA （Salicylic acid） and NAA （1-Naphthalene acetic acid） and stratification on the germination rate of seeds were studied firstly, and then the effects of GA3 and NAA on seedling growth were analyzed. The results showed that the germination rate could be improved by 0.075 g/L SA and stratification markedly. It was indicated by the experiment that the growth of the main root and main stem could be promoted by 0.050 g/L and 0.077 g/L NAA, respectively. However, the leaf length and leaf width decreased significantly in the GA3 and NAA treatments.

Influences of UV-C Irradiation on Seed Germination and Seedling Growth of Winter Wheat

[Objective] This study aimed to investigate the influences of UV-C irradiation on seed germination and seedling growth of winter wheat. [Method] Winter wheat cul- tivar Liaoda 02 was selected as the experimental material and irradiated with UV-C for 1 h,to compare the dynamic changes of seed germination rate under different treatment time and the changes of gibberellin （GA） content and a-amylase activity during the germination process after UV-C irradiation. Wheat seeds irradiated with UV-C for 1 h were sown, the emergence number, growth speed, fluorescence param- eter, reflectance spectrum, relative conductivity and other related physiological indica- tors of seedlings were determined. [Result] The results showed that the germination rate of dry and wet winter wheat seeds irradiated with UV-C for 1 h was improved, compared with the untreated seeds. After soaked for 6, 8, 10 and 12 h, dry seeds irradiated with UV-C for 1 h showed higher GA content and higher a-amylase activity than the control. In addition, UV-C-irradiated dry seeds also showed faster emergence speed and plant growth speed within the first six days after emergence and greater plant height than the control. The flavonoid content in leaves of the treatment group was also increased compared with the control, while the maximal photochemical effi- ciency （Fv/Fm）, photo-chemical reflectance index （PRI）, chlorophyll and carotenoid content were rarely influenced. In conclusion, UV-C irradiation could improve the ger- mination rate of dry seeds and wet seeds soaked for a certain time, especially for wet seeds soaked for 1 h. Furthermore, UV-C-irradiated seeds showed improved ger- mination speed and plant growth. [Conclusion] This study laid the foundation for ex- ploring the effect mechanism of ultraviolet rays on seed generation.

Effect of cold stratification on the temperature range for germination of Pinus koraiensis

Germination at low spring temperatures may offer a competitive advantage for the growth and survival of plant species inhabiting temperate forest ecosystems.Pinus koraiensis is a dominant species in temperate forests of northeastern China.Its seeds exhibit primary morphophysiological dormancy following dispersal in autumn,limiting natural or artificial regeneration:direct seeding and planting seedlings in spring.The aim of this study was to determine the optimum cold stratification temperature that induces germination to increase towards lower temperatures.Seeds from two populations(Changbaishan and Liangshui)were cold stratified at 0,5 and 10℃.Germination to incubation temperatures(10/5,20/10,25/15 and 30/20℃;14/10 h day/night)were determined after 2 and 4 weeks,and 5.5 and6.5 months of cold stratification.After 5.5 months,approximately 68-91%of seeds from both populations germinated at incubation temperatures of 25/15℃and 30/20℃,regardless of cold stratification temperatures.When the cold stratification temperature was reduced to 0℃and the period increased to 6.5 months,germination at 10/5℃significantly improved,reaching 37%and 64%for the Changbaishan and Liangshui populations,respectively.After 6.5 months of cold stratification,there was a significant linear regression between cold stratification temperatures and germination at10/5℃.The range in temperatures allowing for germination gradually expanded to include lower temperatures with decreasing cold stratification temperatures from 10 to 5℃and further to 0℃.