乙烯调控灵芝酸生物合成关键基因的筛选

灵芝是我国著名的药用真菌,灵芝酸是其主要活性成分,具有多种药理活性。乙烯可以促进灵芝酸的生物合成,但其调控机理尚不明确。本实验利用非靶向代谢组研究发现Top 20差异代谢物中含有6种灵芝的活性成分(灵芝酸η、赤芝酸F、赤芝酸N、丹芝酸A、灵芝酸V1和灵芝酸δ),其中有4种灵芝酸(灵芝酸η、赤芝酸F、赤芝酸N和灵芝酸V1)为上调积累,2种灵芝酸(灵芝酸δ和丹芝酸A)为下调积累。通过非靶向代谢组与转录组的关联分析发现基因GL23307、GL25546和GL29595同时与3种灵芝酸积累显著相关,并通过启动子顺式元件预测,发现分别编码泛素蛋白和抑肽酶基因GL25546、GL23307的启动子区域含有响应乙烯信号的顺式作用元件GCC-box,因此,推测这两个基因在乙烯调控灵芝酸生物合成中发挥重要作用。     

烟草水提物对果蝇寿命及有关基因转录的影响

探究烟草的水浸泡液对果蝇寿命与衰老情况的影响以及其中的分子机制。取羽化12 h内的新生W^1118系雄果蝇,分别培养于添加不同浓度烟草浸出液的培养基中处理,每日统计果蝇死亡情况;每7日提取对照组与最高浓度处理组果蝇全RNA,通过Real-time PCR检测过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、去泛素化酶(Rpn11)和去乙酰化酶(Sirt6)等与寿命有关基因的表达水平变化。结果显示,烟草影响使雄果蝇寿命显著缩短,且影响随烟草提取液浓度增加影响逐渐增大;烟草影响下果蝇的抗氧化基因出现显著上调,泛素化调节基因与去乙酰化基因等表达水平的变化均出现显著改变。表明烟草对果蝇的寿命情况有不利影响,可能通过引起氧化胁迫导致果蝇早衰。     

H2S作为下游信号参与SA对低温弱光下黄瓜幼苗光合作用的调控

水杨酸(SA)和硫化氢(H₂S)在调控非生物胁迫下植物生长发育和生理代谢中均起着非常重要的作用,但二者作为信号分子在调控低温弱光下黄瓜光合作用中的互作关系还不清楚。本试验以黄瓜幼苗为试材,分别用SA和硫氢化钠(NaHS,H₂S供体)及其清除剂或抑制剂喷撒叶面,以适宜温光下去离子水处理为对照(CK),研究低温(8 ℃/5 ℃,昼/夜)弱光(100 μmol·m^-2·s^-1)下SA和H₂S对黄瓜幼苗光合作用的调控及互作关系。结果表明: SA可明显增强L-/D-半胱氨酸脱巯基酶(LCD、DCD)活性及其mRNA表达,促进内源H₂S产生;NaHS对苯丙氨酸解氨酶和异分支酸合成酶活性、mRNA表达量及内源SA含量影响不大。SA和NaHS可使低温弱光下黄瓜幼苗的光合速率、气孔导度和蒸腾速率明显提高,胞间CO₂浓度显著降低;同时增强核酮糖-1,5-二磷酸羧化酶、Rubisco活化酶、景天庚酮糖-1,7-二磷酸酯酶和果糖-1,6-二磷酸醛缩酶活性及其mRNA表达,促进光合碳同化;提高光下PSⅡ实际光化学效率和暗下PSⅡ最大光化学效率,从而减轻低温弱光胁迫对黄瓜幼苗的光合机构的损伤和生长量的影响。H₂S清除剂次牛磺酸(HT)可使SA对低温弱光下黄瓜幼苗的光合作用和生长促进效应明显减弱,而SA抑制剂多效唑和氨基茚磷酸对H₂S诱导的黄瓜幼苗光合机构对低温弱光的耐受性无显著影响,说明H₂S作为SA的下游信号,参与调控低温弱光下黄瓜幼苗的光合作用。     

钨酸钠对苹果幼苗15N吸收利用、13C积累及果实品质的影响

分别以1年生苹果砧木M9T337幼苗和5年生烟富3/SH6/平邑甜茶为试材,开展盆栽和田间试验,并结合¹⁵N、¹³C同位素示踪技术,研究不同浓度(0、0.5、1、1.5 mmol·L^-1,分别用CK、T₁、T₂和T₃表示)硝酸还原酶(NR)抑制剂钨酸钠对苹果幼苗¹⁵N吸收利用、¹³C积累和成熟期果实品质的影响。结果表明: 盆栽试验中,喷施0.5~1.0 mmol·L^-1钨酸钠可显著抑制幼苗地上部生长,但对根系生长影响不显著;当钨酸钠浓度达到1.5 mmol·L^-1时可显著抑制根系生长。同一时期各处理叶片NR活性与钨酸钠浓度呈负相关,均表现为CK>T₁>T₂>T₃。随处理时间的延长,叶片硝态氮含量总体表现为先升高后降低的趋势,同一时期各处理硝态氮含量与钨酸钠浓度呈正相关,均表现为T₃>T₂>T₁>CK。喷施钨酸钠可不同程度地降低幼苗各器官¹⁵N吸收量和¹⁵N利用率,且钨酸钠浓度越高,抑制幼苗氮素吸收和利用的效果越显著。随钨酸钠浓度的提高,地上部¹³C积累量呈先升高后降低的趋势,在T₂处理时达到最高;幼苗整株¹³C积累量呈相似的规律。田间试验结果表明,喷施钨酸钠可降低成熟期叶片和果实的氮含量,果皮花青苷含量、果实可溶性固形物、可溶性糖含量和糖酸比均不同程度提高,其中T₂处理的效果最好。综上,T₂处理(1.0 mmol·L^-1钨酸钠)可有效抑制幼苗地上部生长,降低¹⁵N吸收利用,提高¹³C积累,有利于果实品质的提高。     

不同水肥一体化方式对苹果氮素吸收利用特性及产量和品质的影响

以‘嘎啦/八棱海棠’为试材,借助¹⁵N同位素示踪技术,研究了撒施(T₁)、滴灌施氮(T₂)和渗灌施氮(T₃)对嘎啦苹果氮素吸收利用、分配特性和产量品质的影响,以期进一步完善苹果园水肥一体化技术,挖掘提高氮素利用率的途径。结果表明: T₃处理苹果叶片的叶面积、叶绿素和氮含量显著高于T₁和T₂处理。各时期土壤矿化氮(N_min)含量在20~40 cm土层表现为T₃＞T₂＞T₁处理,在0~20 cm土层表现为T₂＞T₃＞T₁处理。同一器官的Ndff值(树体各器官从肥料中吸收到的¹⁵N占该器官全氮量的比例)在各时期均以T₃处理最高,T₂其次,T₁处理最低。果实成熟期的树体¹⁵N利用率表现为T₃＞T₂＞T₁处理,其中T₃处理的树体¹⁵N利用率为24.2%,分别是T₂和T₁处理的1.19和1.65倍。果实成熟期T₁处理的¹⁵N分配率在营养器官最高,T₂处理在贮藏器官最高,T₃处理在生殖器官最高。各处理的单果重、产量、可溶性固形物、硬度、可溶性糖及糖酸比均以T₃处理最高,T₂其次,T₁处理最低。渗灌施氮处理显著促进了嘎啦苹果树体叶片生长和氮素利用,并提高了果实产量和品质。     

水文气候影响下黄河三角洲土壤盐分时空动态

近年来,黄河三角洲在水文气候和人类活动影响下的土壤盐渍化问题日益突出。本研究以东营市河口区、垦利区、东营区和利津县为研究区,选取1985—2018年间20期Landsat系列影像,利用数值回归校正法进行影像光谱一致性转换,在此基础上运用偏最小二乘回归法构建土壤盐分定量反演模型;根据最佳盐分预测模型反演的研究区土壤盐分含量数据,分析土壤盐分(SC)时空变化特征,并探讨水文气候因素的影响。结果表明: 选用10个敏感光谱指数构建的土壤盐分反演模型的预测精度较高,其建模决定系数(R²)、均方根误差(RMSE)为0.769、1.125,验证R²、RMSE和相对分析误差(RPD)分别为0.752、1.203、2.08。利用2016年土壤盐分数据进行反演精度检验,实测值与反演值的R²为0.7279。该模型对1985—2018年20期影像进行土壤盐分反演的结果异常值在10%以内。研究期间,区域平均盐分含量总体呈先上升后下降的趋势,最低的年份为1985年(3.14 g·kg^-1),最高的年份为1995年(5.86 g·kg^-1)。空间变化上,研究区重度盐渍土和盐土面积减少,轻、中度盐渍土面积显著增加(66.6%),盐渍土总面积呈扩大趋势。水文气候条件对土壤盐分的影响具有一定滞后性。气温对土壤盐分积累有促进作用,前半年和前1年的平均气温与含盐量均显著相关,R分别为0.507和0.538;土壤含盐量与区域降水量的相关性不显著,受前季黄河径流量的影响最大,R达-0.543。     

Pythium intermedium,a species complex consisting of three phylogenetic species found in cool-temperate forest ecosystems

Pythium intermedium plays a vital role in the carbon cycle of cool-temperate forests and is widely distributed in Japan's forest soils. In this study, we performed a phylogenetic analysis of the P. intermedium species complex using DNA sequences from multiple loci. The study included 35 isolates from cool-temperate forest soils, seven known P. intermedium isolates, and six known Pythium attrantheridium isolates. We also performed morphological observations and mating tests. Our results showed that all the isolates formed one large clade but were divided into three subclades. Furthermore, we observed many mating reactions between isolates from different subclades, including between P. attrantheridium and P. intermedium. Therefore, we suggest that P. intermedium, P. attrantheridium, and another phylogenetic species belong to one species complex. This is the first report of a species complex within P. intermedium and will be helpful in understanding the evolution of Pythium species in natural ecosystems.     

Isolation and molecular analysis of genes Stpk-V2 and Stpk-V3 homologous to powdery mildew resistance gene Stpk-V in a Dasypyrum villosum accession and its derivatives

Wheat-Dasypyrum villosum translocated chromosomes T6V#2S?6AL and T6V#4S?6DL are known to confer excellent resistance to wheat powdery mildew (PM). However, it is difficult to distinguish the two sources of PM resistance genes through multi-pathotype testing because to date no virulence for them has been found. To reveal the relationship between the PM resistance genes from the two translocations, the sequence of the Stpk-V gene, a key member of powdery mildew resistance locus Pm21, was used as a reference to isolate homologous genes from a D. villosum accession No.1026 and its derivatives 6V#4(6D) disomic substitution (DS) line RW15 and T6V#4S?6DL translocation line Pm97033. Two genes Stpk-V2 and Stpk-V3 were cloned from No.1026. Sequence alignment showed that Stpk-V2 and Stpk-V3 shared 98.2 % and 96.2 % of their DNA and 99.3 % and 100 % of their amino acids in identity with Stpk-V. Compared with Stpk-V, a 22-bp direct sequence repeat and a miniature inverted-repeat transposable element (MITE) were found in the intron 4 of Stpk-V2 and Stpk-V3, respectively. However, Stpk-V2 was not present in DS line RW15 and translocation line Pm97033 based on the PCR result, indicating that Stpk-V2 did not contribute to the PM resistance of RW15 and Pm97033. In the promoter region, a 78-bp insertion was found not only in Stpk-V2 and Stpk-V3, but also in its orthologous gene Stpk-A of wheat. In addition, there was a 17 bp/8 bp deletion/insertion in the putative promoter of Stpk-V3 in comparison with that of Stpk-V/Stpk-V2. Real-time quantitative RT-PCR analysis indicated that the expression levels of Stpk-V and Stpk-V3 genes in the translocation lines were induced by the pathogen, but Stpk-V had a higher expression level than Stpk-V3 at 12 h after inoculation with Bgt. The diversity of Stpk-V gene will help to explore new resistance genes to PM in D. villosum for wheat breeding.