利用CRISPR/CAS9技术编辑水稻香味基因Badh2

【目的】香稻作为一类特殊的水稻群体,以其清香可口的品质特性备受消费者的欢迎。到目前为止,水稻中的香味主要受第8染色体上编码甜菜碱醛脱氢酶基因Badh2控制。【方法】通过CRISPR/CAS9技术对中花11的香味基因Badh2进行编辑。【结果】获得转基因T_0代植株并对其所衍生的T_1代20个单株进行了鉴定分析,获得了一个剔除了载体骨架且第1外显子上插入一个碱基T的突变体材料。该材料中Badh2 RNA水平显著下调;利用GC-MS技术测定野生型及突变体材料籽粒2-乙酰-1-吡咯啉含量,结果表明突变体材料中的香味物质显著增加;此外,我们还对野生型及T_2代香型植株水稻产量及稻米蒸煮食味品质进行了考查及测定分析,发现除分蘖数及结实率呈现出显著差异外(P0.05),其余各项指标在两组材料间都无显著差异。【结论】通过CRISPR/CAS9技术成功地对水稻香味基因进行了编辑,可为香稻育种提供丰富的理论指导,加快香稻的育种进程。     

Dynamics of Bt cotton Cry1Ac protein content under an alternating high temperature regime and effects on nitrogen metabolism

This study was conducted to investigate the effects of alternating high temperature on Cry1Ac protein content on Bt cotton cultivars Sikang 1(SK-1,a conventional cultivar)and Sikang 3(SK-3,a hybrid cultivar).In 2011 and 2012,cotton plants were subjected to high temperature treatments ranging from 32 to 40℃ in climate chambers to investigate the effects of high temperature on boll shell insecticidal protein expression.The experiments showed that significant decline of the boll shell insecticidal protein was detected at temperatures higher than 38℃ after 24 h.Based on the results,the cotton plants were treated with the threshold temperature of 38℃ from 6:00 a.m.to 6:00 p.m.followed by a normal temperature of 27℃ during the remaining night hours(DH/NN)in 2012 and 2013.These treatments were conducted at peak boll growth stage for both cultivars in study periods of 0,4,7,and 10 d.Temperature treatment of 32℃ from 6:00 a.m.to 6:00 p.m.and 27℃ in the remaining hours was set as control.The results showed that,compared with the control,after the DH/NN stress treatment applied for 7 d,the boll shell Cry1Ac protein content level was significantly decreased by 19.1 and 17.5% for SK-1 and by 15.3 and 13.7% for SK-3 in 2012 and 2013,respectively.Further analysis of nitrogen metabolic physiology under DH/NN showed that the soluble protein content and the glutamic pyruvic transaminase(GPT)activities decreased slightly after 4 d,and then decreased sharply after 7 d.The free amino acid content and the protease content increased sharply after 7 d.The changes in SK-1 were greater than those in SK-3.These results suggest that under DH/NN stress,boll shell Cry1Ac protein content decline was delayed.Reduced protein synthesis and increased protein degradation in the boll shell decreased protein content,including Bt protein,which may reduce resistance to the cotton bollworm.     

天山云杉林冠层结构对草本物种多样性的影响

为研究天山云杉林冠层特征与林下草本物种多样性的相互关系，以新疆天格尔森林公园天山云杉林的4个龄级的林分（幼林龄、中林龄、近熟林、成熟林）为研究对象，以冠层分析仪获得的冠层结构特征和林下光环境指数等野外实际调查数据为依据，通过计算 α 多样性指数分析4个龄级的草本物种多样性，利用多元统计方法研究冠层结构对草本多样性的影响。结果表明，各个指标在不同龄级间均有一定的差异，且随着林分的生长和更新，天山云杉林呈现林冠开度逐渐减小、叶面积指数增大、林下光辐射均减弱、物种多样性指数均减小的趋势；典型模型表明林冠开度对冠层结构起到主要作用（呈极显著正相关P<0.01），叶面积指数次之（呈极显著负相关P<0.01）；在林下辐射中，林下总辐射对林内光环境起主要作用；相比冠层结构，林下光照对草本层物种多样性的作用更大，其中林冠开度和丰富度指数之间呈显著正相关，林下总辐射、林下直射辐射和多样性指数、均匀度指数之间均呈显著正相关。     

氮磷减量与液体有机肥配施对膜下滴灌棉田土壤养分的影响

以新疆膜下滴灌棉田为试材,研究氮磷减量与液体有机肥配施对膜下滴灌棉田土壤养分的影响。试验设11个处理:(1)常规施肥(NP);(2)80%常规施肥(80%NP);(3)80%常规施肥+低量氨基酸有机肥(80%NP+LAA);(4)80%常规施肥+中量氨基酸有机肥(80%NP+MAA);(5)80%常规施肥+高量氨基酸有机肥(80%NP+HAA);(6)80%常规施肥+低量黄腐酸有机肥(80%NP+LFA);(7)80%常规施肥+中量黄腐酸有机肥(80%NP+MFA);(8)80%常规施肥+高量黄腐酸有机肥(80%NP+HFA);(9)80%常规施肥+低量沼液有机肥(80%NP+LBS);(10)80%常规施肥+中量沼液有机肥(80%NP+MBS);(11)80%常规施肥+高量沼液有机肥(80%NP+HBS);分别在蕾期、花期、成熟期采集0～20、20～40 cm土层土壤,测定土壤pH值、有机质、氮、磷、钾等有效养分含量。研究结果表明:(1)蕾期时,80%常规施肥配施高量黄腐酸、沼液有机肥显著降低了0～20 cm土层土壤pH值,并对提高土壤速效氮、速效磷含量效果最好。(2)花期时,80%常规施肥配施高量沼液有机肥对提高0～20 cm土层土壤有机质、速效磷含量效果最好。(3)成熟期时,80%常规施肥配施高量沼液有机肥对降低0～20 cm土层土壤pH值、提高土壤有机质含量效果最好。综上所述,新疆滴灌棉田应用80%常规施肥配施高量沼液有机肥效果最好,是新疆滴灌棉田提高土壤肥力和肥料利用率的有效途径。     

裸果木渗透调节物质和抗氧化酶活性对干旱的响应

裸果木(Gymnocarpos przewalskii)起源于古地中海,为荒漠区少有的第三纪孑遗植物种。本文采用2017年6月、7月和8月采集于甘肃安西自然保护区内的裸果木中龄和老龄植株叶片,分析自然干旱影响下叶片渗透调节物质及抗氧化酶活性的变化。结果表明:随着干旱程度的增加,裸果木叶片的可溶性蛋白(SP)显著积累,发挥着重要的渗透调节作用,中龄植株叶片的可溶性糖(SS)含量和老龄植株叶片的脯氨酸(Pro)含量先增加后显著降低,渗透调节作用有限。干旱加剧时,中龄植株叶片的超氧化物歧化酶(SOD)活性和抗坏血酸过氧化物酶(APX)活性呈明显的先增后降。严重干旱时,过氧化物酶(POD)活性和过氧化氢酶(CAT)活性明显增强,说明其通过4种酶的协同作用清除过剩的活性氧,减轻过氧化伤害;老龄植株叶片的CAT活性持续增强,APX活性明显的先升后降,POD活性在重度干旱时显著增强,主要通过这3种酶减轻膜系统伤害。丙二醛(MDA)含量则随干旱程度增大显著增加,膜脂过氧化程度不断加深。在相同的降雨条件下,裸果木中、老龄植株的Pro、SS、SP含量、POD和CAT活性、MDA含量对干旱的响应程度存在显著差异,隶属度函数评价法表明,老龄植株的抗旱能力大于中龄植株,表明裸果木的抗旱性随年龄的增大而提高。     

基于转录组测序的半滑舌鳎长链非编码RNA的初步鉴定与分析

以抗病家系与易感家系半滑舌鳎为材料,进行哈维弧菌感染实验,并对易感家系感染前(CsSU)、易感家系感染后(CsSC)、抗病家系感染前(CsRU)、抗病家系感染后(CsRC)4组进行转录组测序,根据RNA-seq数据挖掘半滑舌鳎长链非编码RNA信息;通过生物信息学分析,筛选出与抗哈维弧菌病相关的差异长链非编码RNA(long non-coding RNA, lncRNA)。结果显示,共识别出4 584个lncRNA座位,包含5 714个转录本;其基本特征与编码基因的比较分析,lncRNA的GC含量低于编码基因,单外显子基因数多于编码基因,转录本的平均长度长于编码基因,基因表达量低于编码基因。对4组样品进行两两比较(CsRU vs CsSU、CsRC vs CsSC、CsRC vs CsRU、CsSC vs CsSU)分别筛选出818、813、261、140个差异表达lncRNA,其中CsRU与CsSU之间、CsRC与CsSC之间lncRNA数目差异最多,通过聚类分析确定了各实验组的表达模式之间的联系,CsRU与CsSU之间的表达模式最为相近。通过共表达分析,预测出lncRNA和274个编码基因可能存在14 539种相互关系,并进行了功能注释,进而筛选出7个关键lncRNA。qRT-PCR结果显示,差异表达lncRNAs的表达模式和转录组数据得到的基本一致。研究结果为揭示lncRNA在半滑舌鳎抗哈维弧菌免疫调控反应中的作用提供重要的参考数据。     

One-time fertilization at first flowering improves lint yield and dry matter partitioning in late planted short-season cotton

Cotton producers have substantially reduced their inputs(labor, nutrients, and management) mainly by adopting a shortseason cropping management that is characterized by late sowing, high density, and reduced fertilization with one-time application at the first bloom stage without lint yield reduction. However, it has been hypothesized that one-time fertilization at an earlier growth stage could be a more effective and economic management practice. A two-year field experiment was conducted by applying five fertilizer one-time fertilization at 0(FT1), 5(FT2), 10(FT3), 15(FT4), and 20(FT5) days after the first flower appeared in the field and one three-split fertilizer application taken as the conventional control(FT6), making six treatments altogether. Cotton growth period, biomass accumulation, yield, and its formation were quantified. The results showed that the one-time fertilization did not affect the cotton growth progress as compared to FT6, however, the total crop cycles for FT3–FT5 were 3 days shorter. FT1 produced the highest cotton lint yield(1 396 kg ha–1), which was similar to the FT6 but higher than the other treatments, and could be attributed to more bolls per unit area and higher lint percentage. Cotton yield was positively correlated with cotton plant biomass accumulated. FT1 had both the highest average(VT)(193.7 kg ha–1 d–1) and the highest maximum(VM)(220.9 kg ha–1 d–1) rates during the fast biomass accumulation period. These results suggest that one-time fertilizer application at the first flower stage might be an adjustment that is more effective than at first bloom, and allowed for easier decision making for application date due to non counting of plants with flowers is needed.     

Detection of seven phytohormones in peanut tissues by ultra-highperformance liquid chromatography-triple quadrupole tandem mass spectrometry

Development of highly sensitive and reliable method for detection of phytohormones is of great significance to study plant hormones and agricultural production.In this study,an ultra-high-performance liquid chromatography-mass spectrometry/mass spectrometry method was established for separation and quantification of trans-zeatin,trans-zeatin riboside,gibberellin A_3,indol-3-acetic acid,salicylic acid,abscisic acid,and jasmonic acid(JA) without any label.The sepa ration was performed on an Agilent Explus Plus C18 column by using methanol and water as mobile phases with gradient elution.The target compounds were confirmed and quantified by mass spectrum via positive electrospray ionization for trans-zeatin,transzeatin riboside,indole-3-acetic acid,and via negative electrospray ionization for gibberellin3,salicylic acid,abscisic acid,and JA.The limits of detection ranged from 0.0127 ng L~(-1) for gibberellin A_3(GA_3) to 33.26 ng L~(-1) for JA and were lower than the currently reported values in literature.The proposed method was applied for qualitative and quantitative analyses of phytohormones in peanut gynophores and pods.The recoveries of the spiked phytohormones ranged from 80.20 to102.56%.The contents of seven endogenous hormones varied specifically in different development stages of peanuts.This study provides a highly sensitive and selective detection method for hormones and elucidates the growth and development of the gynophore and peanut fruit,which are controlled by seven endogenous hormones.