不同提取方法对红松籽油提取效果及功能性质的影响

  目的  立足于溶剂法,通过对比超声波、微波、光波单独处理较短时间的红松籽油的得率,以及处理后单独浸提红松籽油得率的增加情况,明确在红松籽油提取过程中处于主导作用的方法,同时确认4种提取方法对红松籽油的理化指标、脂肪酸和抗氧化能力的影响。  方法  按照GB/T 5009.229—2016《食品中酸价的测定》、GB/T 5532—2008《动植物油脂 碘值的测定》和GB/T 5009.227—2016《食品中过氧化值的测定》分别测定4种方法提取的红松籽油的酸值、碘值和过氧化物值,采用气相色谱–质谱联用法测定红松籽油中脂肪酸的种类和质量分数,采用分光光度法测定红松籽油对DPPH·和ABTS+·的清除能力。  结果  正己烷作为最佳提取溶剂,当提取温度25 ℃,液料比18 mL/g,浸提时间5 h红松籽油得率为65.52%。超声波法（300 W,10 min）、微波法（380 W,100 s）、光波法（400 W,11 min）单独处理时的红松籽油得率分别为67.63%、62.26%、58.25%。上述3种方法处理后再单独使用正己烷浸提,使总提取时间达到5 h后的红松籽油得率分别增加了2.18%、4.50%、6.10%。4种方法提取的红松籽油符合GB/T 2716—2018《食品安全国家标准 植物油》中酸值（≤ 4 mg/g）、过氧化值（≤ 0.25 g/100 g）的限量标准。溶剂法制备出的红松籽油的酸值和过氧化值均最低（P < 0.05）。脂肪酸质量分数由高到低分别是亚油酸（44.54% ~ 46.32%）、油酸（28.29 % ~ 28.83%）和皮诺敛酸（13.15% ~ 14.51%）。溶剂法、超声波法、微波法、光波法所提取的红松籽油清除DPPH·和ABTS+·的IC₅₀值分别是9.41、8.80、9.43、9.61 g/L和5.10、5.51、6.10、5.43 g/L。  结论  对超声波法、微波法和光波法单独作用及浸提后得率进行比较,发现超声波法、微波法和光波法在提取过程中起主导作用。与溶剂法、微波法和光波法相比,超声波法提取的红松籽油得率高,不饱和脂肪酸质量分数高,抗氧化能力强,能够达到较好的提取效果,此方法应用于红松籽制油工业中具有一定优势。      

Expression profiling of transgenes (Cry1Ac and Cry2A) in cotton genotypes under different genetic backgrounds

Transgenic cotton carrying the Cry1Ac gene has revolutionized insect pest control since its adoption, although the development of resistance in insect pests has reduced its efficacy.  After 10 years of cultivating Bacillus thuringiensis (Bt) cotton with a single Cry1Ac gene, growers are on the verge of adopting Bt cotton that carries the double gene (Cry1Ac+Cry2A) due to its better effectiveness against insect pests.  Thus, the current study was designed to evaluate the role of each gene in the effectiveness of Bt cotton carrying the double gene.  The expression levels of the Cry1Ac and Cry2A genes were evaluated in the leaves of 10 genotypes (2 parents and 8 F₁ hybrids) at 30 days after sowing (DAS), while samples of leaves, bolls and flowers were taken from the upper and lower canopies at 70 and 110 DAS.  The F₁ hybrids were developed through reciprocal crosses between two Bt (CKC-1, CKC-2) and two non-Bt (MNH-786, FH-942) parents.  The differential expression of transgenes was evaluated through Enzyme Linked Immuno-Sorbent Assay (ELISA).  The results showed that the MNH786×CKC-1 hybrid had the highest concentrations of Cry1Ac gene at 30 DAS (3.08 µg g^–1) and 110 DAS (1.01 µg g^–1) in leaves.  In contrast, the CKC-2×MNH-786 hybrid showed the lowest concentrations of Cry1Ac gene at 30 DAS (2.30 µg g^–1) and 110 DAS (0.86 µg g^–1).  The F₁ hybrid FH-942×CKC-2 showed the highest concentrations of Cry2A gene at 30 DAS (8.39 µg g^–1) and 110 DAS (7.74 µg g^–1) in leaves, while the CKC-1×MNH-786 hybrid expressed the lowest concentrations of Cry2A gene at 30 DAS (7.10 µg g^–1) and 110 DAS (8.31 µg g^–1).  A comparison between the two stages of plant growth showed that leaves had the highest concentrations at 30 DAS, whereas the lowest concentrations were observed at 110 DAS for both genes in leaves.  When the expression pattern was compared between various plant parts in genotype CKC-2, it was found that leaves had higher concentrations of Cry1Ac (3.12 µg g^–1) and Cry2A (8.31 µg g^–1) at 70 DAS, followed by bolls (Cry1Ac (1.66 µg g^–1) and Cry2A (8.15 µg g^–1)) and flowers (Cry1Ac (1.07 µg g^–1) and Cry2A (7.99 µg g^–1)).  The genotype CKC-2 had higher concentrations of Cry1Ac (3.12 µg g^–1) and Cry2A (8.31 µg g^–1) in the upper canopy but less accumulation (2.66 µg g^–1 of Cry1Ac, 8.09 µg g^–1 of Cry2A) in the lower canopy at 70 DAS.  Similarly, at 110 DAS, the expression levels of Cry1Ac and Cry2A in upper and lower canopy leaves were 1.52 and 7.92 µg g^–1, and 0.99 and 7.54 µg g^–1, respectively.  Hence, the current study demonstrates that different genotypes showed variable expression for both of the Cry1Ac and Cry2A genes during plant growth due to different genetic backgrounds.  The Cry2A gene had three-fold higher expression than Cry1Ac with significant differences in expression in different plant parts.  The findings of this study will be helpful for breeding insect-resistant double-gene genotypes with better gene expression levels of Cry1Ac and Cry2A for sustainable cotton production worldwide.     

Predictive models of drought tolerance indices based on physiological,morphological and biochemical markers for the selection of cotton (Gossypium hirsutum L.) varieties

The use of tolerant crop varieties is a strategy that mitigates the water deficit effect in a sustainable way. The generation of these varieties is more efficient when variables associated with this tolerance have been identified, since they can facilitate the breeding processes. This study aimed to establish the relationships between water deficit tolerance of four cotton varieties (Nevada-123, Oasis-129, Guatapuri, and Festivalle) and morphological variables (monopodial branches, boll weight, root/shoot ratio, and leaf and root dry matter), physiological variables (relative water content, net photosynthesis, stomatal conductance, electron transport rate, photochemical quenching, photochemical efficiency of PSII, chlorophyll a/b ratio (Chl a/b), C¹²/C¹³ isotope ratio, and electrolyte leakage), and biochemical variables (contents of sugars, proline, carotenoids, and malondialdehyde). Furthermore, calibrated predictive models of the drought tolerance indices were developed based on the key variables identified. For this purpose, a pot experiment was established where plants were subjected to a moderate or severe water deficit during the blooming stage for 12 days. The stress tolerance index (STI) and mean productivity (MP) were calculated. For the evaluated variables, the differences between well-watered and water deficit plants (Δ) were calculated and ANOVA, partial least squares, Pearson's correlation, and multiple linear regression analyzes were performed. A model was generated that explained 95% of the STI and was composed of Δmalondialdehyde, Δproline, and Δboll weight. For MP, the model was comprised of Δstomatal conductance, Δroot/shoot ratio, and ΔChl a/b, and explained 89% of the MP. The analysis of the assessed variables allowed the identification of key variables and the development of calibrated predictive models that can be used in screening to obtain cotton varieties with different levels of water deficit tolerance.     

Tomato mottle mosaic virus: Characterization,resistance gene effectiveness,and quintuplex RT-PCR detection system

Tomato mottle mosaic virus (ToMMV), an economically important species of the genus Tobamovirus, causes significant loss in yield and quality of tomato fruits. Here, we identified the Shandong isolate of ToMMV (ToMMV-SD) collected from symptomatic tomato fruits in Weifang, Shandong Province of China. ToMMV-SD caused symptoms such as severe mosaic, mottling, and necrosis of tomato leaves, yellow spot and necrotic lesions on tomato fruits. The obtained full genome of ToMMV-SD was 6 399 nucleotides (accession number MW373515) and had the highest identity of 99.5% with that of isolate SC13-051 from the United States of America at the genomic level. The infectious clone of ToMMV-SD was constructed and induced clear mosaic and necrotic symptoms onto Nicotiana benthamiana leaves. Several commercial tomato cultivars, harboring Tm-2² resistance gene, and pepper cultivars, containing L resistance gene, were susceptible to ToMMV-SD. Plants of Solanum melongena (eggplant) and Brassica pekinensis (napa cabbage) showed mottling symptoms, while N. tabacum cv. Zhongyan 100 displayed latent infection. ToMMV-SD did not infect plants of N. tabacum cv. Xanthi NN, Brassica rapa ssp. chinensis (bok choy), Raphanus sativus (radish), Vigna unguiculata cv. Yuanzhong 28-2 (cowpea), or Tm-2² transgenic N. benthamiana. A quintuplex RT-PCR system differentiated ToMMV from tomato mosaic virus, tomato brown rugose fruit virus, tobacco mosaic virus, and tomato spotted wilt virus, with the threshold amount of 0.02 pg. These results highlight the threat posed by ToMMV to tomato and pepper cultivation and offer an efficient detection system for the simultaneous detection of four tobamoviruses and tomato spotted wilt virus infecting tomato plants in the field.     

Heterosis and heterotic patterns of maize germplasm revealed by a multiple-hybrid population under well-watered and drought-stressed conditions

Understanding the heterosis in multiple environments between different heterotic groups is of fundamental importance in successful maize breeding. A total of 737 hybrids derived from 41 maize inbreds were evaluated over two years, with the aim of assessing the genetic diversity and their performance between heterotic groups under drought-stressed (DS) and well-watered (WW) treatments. A total of 38 737 SNPs were employed to assess the genetic diversity. The genetic distance (GD) between the parents ranged from 0.05 to 0.74, and the 41 inbreds were classified into five heterotic groups. According to the hybrid performance (high yield and early maturity between heterotic groups), the heterosis and heterotic patterns of Iowa Stiff Stalk Synthetic (BSSS)×Non-Stiff Stalk (NSS), NSS×Sipingtou (SPT) and BSSS×SPT were identified to be useful options in China's maize breeding. The relative importance of general and specific combining abilities (GCA and SCA) suggests the importance of the additive genetic effects for grain yield traits under the WW treatment, but the non-additive effects under the DS treatment. At least one of the parental lines with drought tolerance and a high GCA effect would be required to achieve the ideal hybrid performance under drought conditions. GD showed a positive correlation with yield and yield heterosis in within-group hybrids over a certain range of GD. The present investigation suggests that the heterosis is due to the combined accumulation of superior genes/alleles in parents and the optimal genetic distance between parents, and that yield heterosis under DS treatment was mainly determined by the non-additive effects.     

5%调环酸钙EA对小麦抗倒伏和产量及其相关因素的影响

为了探讨调环酸钙对小麦产量相关因素及安全性的影响,以小麦品种‘豫麦49’为材料,返青后施用不同剂量的5%调环酸钙EA开展田间试验。结果表明,小麦返青期施用5%调环酸钙EA 150~ 600 g/hm²,可矮化植株高度,降低基部节间长度、增加茎粗和茎壁厚度。调环酸钙能提高小麦穗数、穗粒数、千粒重和产量。返青期施用5%调环酸钙EA 300 g/hm²和5%调环酸钙EA 300 g/hm²+0.01%芸苔素内酯SL 150 mL/hm²的产量较高,分别达7410.0 kg/hm²和7560.0 kg/hm²,较对照(CK)分别增产10.7%和13.0%。5%调环酸钙EA对小麦安全无药害,建议在小麦上推广使用。     

田间盆栽接种法鉴定若干向日葵品种(系)列当抗性

向日葵列当( Orobanche Cumana Wallr.) 是一种寄生性杂草,给向日葵生产造成严重威胁。为鉴定向日葵抗列当品种并为抗列当育种工作提供优异的种质资源,利用田间盆栽接种法鉴定向日葵新杂交组合以及恢复系、不育系对向日葵列当的抗性。于2019年对112个向日葵新品种(系),2020年对168个向日葵新品种(系)分别进行列当抗性鉴定。结果显示：2019年筛选出16个免疫,5个高抗的杂交组合;24个免疫、2个高抗的恢复系;7个免疫、2个高抗的不育系。2020年筛选出35个免疫、9个高抗的杂交组合;38个免疫,25个高抗的恢复系。2020年向日葵材料鉴定结果以免疫、高抗、中抗等抗性型为主,且均未发现高感材料,主要由于无论是杂交组合或恢复系,其亲本均来自之前几年鉴定得到抗性较好的种质资源。本单位经多年盆栽接种鉴定及自然发生列当的大田抗性验证,已获得若干抗性稳定的向日葵食葵、油葵品种(系)。     

混合盐胁迫对白榆幼苗形态及生理指标的影响

为揭示混合盐胁迫对白榆幼苗形态及生理指标的影响,以白榆幼苗为试验材料,采用盆栽法,设置6个混合盐（等浓度NaCl和NaHCO₃溶液）浓度［0（CK）、10（T1）、30（T2）、50（T3）、70（T4）、100（T5） mmol·L^-1］对白榆幼苗进行不同时间胁迫处理,分析混合盐胁迫下白榆幼苗的生长及生理指标的变化。结果表明,白榆幼苗生长对盐胁迫反应敏感,当盐浓度（10 mmol·L^-1）较低时即表现出明显的抑制作用,胁迫35 d后,随着盐浓度的升高,白榆幼苗的总生物量、株高及地径增长量均显著降低,当混合盐浓度高于50 mmol·L^-1时,各处理间降幅趋于稳定。不同盐浓度胁迫下,根冠比值均大于对照组;随着盐胁迫程度的加剧,丙二醛含量和相对电导率在不同处理时间下均逐渐增大,而可溶性糖、过氧化物酶及超氧化物歧化酶均呈现先升高后降低的趋势;不同处理时间下,叶绿素含量随着盐胁迫浓度的增加显著降低,混合盐浓度为100 mmol·L^-1时,不同处理时间下叶绿素含量均为最小值。混合盐胁迫对白榆幼苗的生长具有一定程度的抑制作用,但白榆幼苗可通过渗透调节及提高保护酶活性主动适应逆境,从而表现出较强的抗盐能力。研究结果旨在为白榆在盐碱地区的种植提供理论依据和技术参考。     

陆地棉花铃期抗旱指标筛选及评价

为分析棉花资源的抗旱性,以253份陆地棉资源为试验材料,于花铃期进行干旱胁迫处理,测定包括光合作用、农艺性状和产量性状相关的15个指标,通过主成分分析、相关性分析、抗旱隶属函数分析及聚类分析,对其进行抗旱性综合评价及抗旱指标筛选。以抗旱性度量值（D值）为参数,可将上述资源分为强抗旱、抗旱、耐旱、较敏感、敏旱5类。其中,强抗旱材料包括JK625、库车7946、肯尼亚2号等14份;敏旱材料包括HT-176、川R128、鄂棉18号等19份。有效铃数、气孔导度、蒸腾速率、叶绿素相对值、有效果枝数、皮棉重和株高较其他指标更为敏感,可作为花铃期抗旱性鉴定的关键指标;另外,叶绿素相对值、株高、气孔导度和蒸腾速率可作为花铃期抗旱的立测指标。     

44份青海小麦品种（系）抗条锈性评价及抗条锈病基因检测

【目的】探明青海主栽与后备小麦品种（系）对小麦条锈病的抗性水平及部分抗条锈病基因的分布,为青海省高抗条锈病小麦品种的选育提供理论基础。【方法】利用小麦条锈菌优势生理小种CYR31、CYR32、CYR33、CYR34对44份供试小麦品种分别在温室和田间进行全生育期和成株期抗病性鉴定,利用抗条锈病基因Yr5、Yr9、Yr10、Yr15、Yr24、Yr44的侧翼分子标记检测其抗病基因的分布。【结果】在供试的44个小麦品种（系）中,青麦1号、青春533和青春38等31份（70.4％）材料对CYR31表现抗病,青麦1号、青春533和青春38等35份（79.5％）材料对CYR32表现抗病,青麦1号、青春38、YZ282等35份（79.5％）材料对CYR33表现抗病,互麦13、云繁105、稷麦8号等35份（79.5％）材料对CYR34表现抗病;青春533、青春38、青春39等20份材料（系）表现成株期抗病性,占供试品种的45.4％,其中GY363和GY856 表现免疫,15-34-3、HYNM19-2和互紫麦1号等11份小麦品种表现高抗,青春38、青春39和稷麦8号等7份小麦品种（系）表现中抗,分别占总数的4.5％,25.0％和15.9％。分子检测结果表明,15-34-3、HYNM19-2、GY363等5份（11.3％）材料可能携带Yr5基因,云繁105、稷麦8号和15-34-3等11份（25.0%）材料可能携带Yr9基因,青春533、青春38、40-12-86等4份（9.1％）材料可能携带Yr10基因,互紫麦1号、YGW-46、GY363等5份（11.3％）材料可能携带Yr15基因,青麦5号、稷麦8号及15-34-3等15份（34.1％）材料可能携带Yr24基因,宁春51、15-34-3、19399-11等8份（18.1％）材料可能携带Yr44基因,其中航远L621未携带这6个抗病基因,但是全生育期表现抗病。【结论】青海省部分小麦品种（系）抗病性较高,今后应减少含有Yr9基因小麦品种的使用,加大含有多个抗病基因组合品种选育的力度。