黑松（Pinus thunbergii Parl.）是暖温带海岸防护林的建设中的先锋树种，我国将其从日本引种到盐碱地区进行生态修复。利用外生菌根菌与黑松合成菌根化苗，是提高黑松耐盐碱性和在盐渍土壤造林成活率的一项重要措施。目前关于黑松的菌根合成研究较少。本文研究了实验室分离保存的18种菌根真菌菌株在盐碱胁迫条件下的培养特性，筛选、优化了黑松菌根化苗合成基质，成功合成了14种黑松菌根化苗，并研究了黑松菌根苗对盐碱胁迫的生理响应，结果如下： 1、所选用的18种外生菌根真菌菌株为：土生空团菌（Cenococcum geophilum，编号CG）、红汁乳菇（Lactarius hatsudake，编号LH）、紫晶蜡蘑（Laccaria amethystea，编号LA）、彩色豆马勃（Pisolithus tinctorius，编号PT）、须腹菌（Rhizopogon superiorensis，编号RS）、马勃状硬皮马勃（Scleroderma areolatum，编号SA）、褐环粘盖牛肝菌（Suillus luteus）的两个菌株分别编号SL1和SL2、Melanogaster intermedius编号MI，及待鉴定9种菌株分别编号为LAC、UJ、U1、U9、U10、U11、U12、U14、U15。在供试菌株中只有CG菌同时具有耐盐性和耐碱性。CG、LAC、U10、UJ、MI、SL2、U9、U1、RS、SA、LAJ菌有耐盐胁迫能力，根据其相对生长率判断其耐盐胁迫能力大小依次为CG、LAC > U10 > UJ 》MI、SL2、U9 > U1、RS、SA 》LAJ（“》”表示在P<0.05条件下其左侧显著大于右侧，“>”表示在P<0.05条件下其左侧大于右侧但是差异不显著，下文同）。MI、U14、CG、SL2、LAJ、LH、UJ、U9菌有耐碱胁迫能力，其耐碱性依次为MI、U14》CG》SL2、LAJ、LH》UJ》U9。CG、MI、SL2、LH、LAJ、U14菌在碱处理条件下液体培养基pH值都有显著下降（P<0.05）。 2、以火山灰、水草、椰壳、树皮、兰花石、陶粒、珍珠岩、蛭石、石英砂为材料，根据不同配比配成26种不同基质。通过菌根苗合成试验筛选发现在实验室条件下火山灰最适宜用来做接菌基质，其理化性质为：pH:5.7-6.5，EC:0.02-0.03 dS·m-1，容重：0.28-0.54 kg·m-3， 持水能力：190%-305%，总孔隙度：72%-108%，气水比：0.27-1.40。 3、利用本实验设计的菌根合成基质成功合成14种菌根化苗，其侵染时间在2个月左右。除SL1侵染率为15.4%外，其余菌根化苗侵染率达到60%以上。除CG、LA、RS、SL1、SL2外，其余菌尚未在野外调查中发现与黑松共生，同时也是第一次在实验室条件下人工合成菌根化苗。与非接种相比，接种外生菌根真菌对黑松有明显的促生效果，接菌后植株的生物量、地径、株高、茎长、根长有不同程度的增长，其中PT、U9、LAJ、TM、LAC、SL1、U11、RS、CG、U10、UZ菌根化苗与对照相比生物量显著增加（p<0.05），分别增加了20.5%、30.1%、34.4%、45.1%、51.9%、60.3%、66.1%、94.7%、126.9%、241.2%。LAJ、U10、CG、UZ菌的菌根化苗地径比对照分别提高了64.5%、64.8%，77.2%、82.1%达到极显著水平（p<0.01）。接LAJ、CG、U11、SL1、TM后植株的茎长与不接菌相比有显著增长（p<0.05），分别增加了29.2%、26.9%、27.7%、29.4%、29.3%。根系空间吸收面积大大增加，菌根形态各异，其菌丝体颜色，分支方式及根毛有无等菌根根尖形态方面存在不同或相似特征。 4、通过土培试验，研究了LAJ、PT、CG三种菌根化黑松幼苗的耐盐碱性。结果表明，盐渍土培2个月对CG、LAJ菌根化苗的正常生长几乎无影响，而PT菌根化苗与非菌根化苗则出现叶片萎黄甚至死亡等现象。与非菌根化苗相比，菌根化苗的叶绿素a和叶绿素b和类胡萝卜素的含量显著高于与非菌根化苗，地下部的K/Na含量均有显著提高。CG、LAJ菌根化苗的叶绿素a、b含量和体内脯氨酸的含量均显著高于PT菌根化苗（P<0.05）。LAJ、PT菌根化苗具有一定的耐盐碱能力，有望用于滨海盐渍土的造林实践。

Pinus thunbergii Parl. is a pioneer specie in the construction of temperate coastal shelterbelt, and it was used in biorestoration of saline/alkaline areas and was introduced from Japan. It is an important measure to improve the salinity tolerance and afforestation of Pinus thunbergii Parl. in saline soils by using the ectomycorrhizal Pinus thunbergii Parl.. However, less attention was paied in the study of synthesis of the Pinus thunbergii Parl. ectomycorrhizas. In this paper, 18 of mycorrhizal fungi strains were used to compare their saline/alkaline-tolerance; the synthetic substrate of black pine mycorrhizal seedlings was optimized; and 14 of ectomycorrhizal pine seedlings were successfully synthesized. The physiological responses of ectomycorrhizal pine seedlings on saline stress were also studied, the results are as follows: 1, A total of 18 ectomycorrhizal fungi strains was used in this study, including Cenococcum geophilum, No.CG, Lactarius hatsudake, No.LH, Laccaria amethystea, No.LA, Pisolithus tinctorius, No.PT, Rhizopogon superiorensis, No.RS, Scleroderma areolatum, No.SA, Suillus luteus: the two strains was No.SL1 and SL2, Hysterangium sp., No.U14, Ileodictyon gracile, No.U15, Melanogaster intermedius, No.MI, and 9 unidentified strains signed UJ, U1, U9, U10, U11, respectively. Only CG has both salt and alkali resistance among the tested strains. CG, LAC, U10, UJ, MI, SL2, U9, U1, RS, SA, LAJ can tolerate salt stress. Based on their relative growth rate, their ability to salt stress is CG, LAC> U10> UJ "MI, SL2, U9> U1, RS, SA "LAJ (" "》" indicates P <0.05 condition was significantly greater than the right to the left, ">" indicates P <0.05 greater than the right to the left under the conditions but the difference was not significant , hereinafter the same). MI, U14, CG, SL2, LAJ, LH, UJ, U9 strain have the alkali stress capacity (MI、U14》CG》SL2、LAJ、LH》UJ》U9). 2, By screening of 26 different substrates in laboratory conditions, volcanic ash is the most suitable substrates used for inoculation, and its physical and chemical properties as: pH :5.7-6.5, EC :0.02-0.03 dS · m-1, bulk density :0.28-0.54 kg/m3, water holding capacity: 190.13% -304.74%, total porosity: 72.30% -107.69%, gas-water ratio: 0.27-1.40. 3, A total of 14 ectomycorrhizal seedlings were successfully synthesized using the optimized synthetic substrate. The infection time was within two months, and the infection rate can reach 60% or more, except 15.4 % of the SL1-mycorrhizal seedlings. In addition to CG, LA, RS, SL1, SL2, the remaining strains have not been found in the field survey symbiosis with black pine, and it is the first time to obtained synthetic mycorrhizal seedlings under laboratory conditions. Compared with non-inoculation, the ectomycorrhizal fungi have significant growth-promoting effect of the host plant in the biomass, diameter, plant height, stem length, root length. For example, the biomass of mycorrhizal seedlings infected by PT, U9, LAJ, TM, LAC, SL1, U11, RS, CG, U10, UZ was significantly (p <0.05) increased of 20.5%, 30.1%, 34.4%, 45.1 %, 51.9%, 60.3%, 66.1%, 94.7%, 126.9%, 241.2%, respectively, compared with the control seedlings. Root space absorption area was greatly increased in mycorrhizal seedlings. There are different or similar features in the ectomycorrhizal shapes, mycelium color, the amount of lateral roots and root hairs, as well as the morphology of mycorrhizal root tips.. 4, Soil pot experiment was arranged to study the physiological responses of LAJ-, PT-, CG-mycorrhizal pine seedlings to saline/alkaline stress. The results showed that there was no apparent toxic effects of salt stress in CG-, LAJ-mycorrhizal seedlings two months after soil culture, while PT-mycorrhizal and non-mycorrhizal seedlings showed leaf chlorosis, even death phenomena. Compared with non-mycorrhizal seedlings, chlorophyll a and chlorophyll b and carotenoid content of mycorrhizal seedlings was significantly higher than those of non-mycorrhizal seedlings. The ratio of K/Na in roots were significantly increased in mycorrhizal seedlings. Among three fungi species, CG, LAJ led higher chlorophyll a, b content and proline levels in host plants compare with PT (P <0.05). The results showed that LAJ- and PT-mycorrhizal seedlings had a certain salinity tolerance capacity, and could be used in the afforestation of coastal saline soils.