Molecular identification of sex in Hippophae rhamnoides L. using isozyme and RAPD markers

In many dioecious plants, gender affects economic value, breeding schemes and opportunities for commercial harvests. Hippophae rhamnoides L. is a dioecious plant species in which female genotypes are commercially preferred over male genotypes. Its berries have rich medicinal, nutritional and pharmaceutical properties because of their large amounts of vitamins, essential oils, proteins, fatty acids, free amino acids and flavanoids. Primary limitation for breeding H. rhamnoides L. is its dioecious nature, since gender cannot be identified by traditional methods. Therefore, some reliable and quick methods need to be developed. This commu- nication deals with the development of isozyme and RAPD markers for early sex identification in this dioecious tree. The isozyme analysis was conducted with four enzyme systems, viz. peroxidase, esterase, malate dehydrogenase and catalase. The peroxidase enzyme system produced a female specific sex marker, which successfully differentiated between the staminate and pistillate geno- types ofH. rhamnoides L. Thirty five random decamer primers were used in our study and one male sex linked marker was identified. OPD-20 （5＇-ACTTCGCCAC-3＇） displayed a band at 911 bp that expressed polymorphism between male and female genotypes. The staminate and pistillate genotypes could be distinguished using RAPD marker OPD-209n. These results revealed the immense poten- tial of peroxidase isozyme patterns and RAPD as genetic markers for sex identification in H. rhamnoides L.

Analysis of Effective Composition of Hippophae Rhamnoides L.

The vitamin E, proteins, amino acids, sugar, trace elements and fatty acids in the oil of Hippophae Rhamnoides L. seeds were quantitatively analyzed. Several methods for analyzing vitamin E were compared with each other and the content of gossypol in the oil was first determined.

Short-term responses to salinity of seabuckthorn (Hippophae rhamnoides L.) seedlings in the extremely cold and saline Qinghai region of China

The investigation of the mechanisms of plant adaptation to stressor action is one of the leading directions of current biological studies. To understand the mechanism of salt tolerance of seabuckthom （Hippophae rhamnoides L.） and identify its ability to cope with the salinity effect in the saline and extremely cold region of Qinghai, China, a test was conducted with two-year-old seedlings subjected to 0, 200, 400 and 600 mmol·L^-1 NaClsolutions for 30 d. The results show that with an increase in salinity, the biomass of H. rhamnoides seedlings clearly decreased. Leaf water potential （ψw） and relative water content （RWC） were significantly reduced under salinity, with severe water shortages appearing in leaves. At the same time, the total chlorophyll content declined markedly. When salinity increased and stress time prolonged, the net CO2 assimilation rate （A） significantly declined. Intercellular CO2 concentration （Ci） declined at first and was then followed by an increase over the stress time. We conclude that H. rhamnoides grown in the extremely cold and saline region of Qinghai has a certain resistance to salt, which can be planted at appropriate salinity levels.

Effects of Total Flavonoids ofHippophae RhamnoidesL.on Intracellular Free Calciumin Cultured Vascular Smooth Muscle Cells of Spontaneously Hypertensive Rats and Wistar-Kyoto Rats

To explore the effects of total flavonoids of Hippophae rhamnoides L. （TFH） quercetin （Que） and isorhamnetin （Isor） on the intracellular free calcium （[Ca^2＋]） in vascular smooth muscle cells （VSMC） of spontaneously hypertensive rats （SHR） and Wistar-Kyoto rats （WKY）. Metheds： Fluo 3-acetoxymethylester（Fluo-3/AM） was used to observe the effects of TFH （100mg/L） and its essential monomers, namely Que （10^-4mol/L） and Isor （10^-4mol/L） on changes of [Ca^2＋]1 in cultured SHR and WKY VSMC （abbr. to Ca-SHR ＆ Ca-WKY） following exposure to high K^＋, norepinephrine （NE） and angiotensin Ⅱ （AngⅡ）, and to compare with the effects of verapamil （Ver）. Results： （1） TFH, Que and Isor had inhibitory effects on resting Ca-SHR （P〈0.05）, but had no significant effects on Ca-WKY （P〉0.05）. （2） High K^＋ could increase Ca-SHR more significantly than Ca-WKY （P〈0.05）; TFH, Que and Isor could inhibit the elevation of [Ca^2＋]1 induced by high K^＋ -depolarization, with the effects similar to that of Ver, and the effect on Ca-SHR was more significant than that on Ca-WKY （P〈0.05）. （3） NE and Ang Ⅱ could increase Ca-SHR more significantly than Ca-WKY （P〈0.05）, TFH, Que and Isor had remarkably inhibitory effect on the elevation of Ca-SHR and Ca-WKY induced by NE or Ang Ⅱ. （4） In the absence of extracellular Ca^2＋ , TFH, Que and Isor also had certain inhibitory effect on Ca-SHR and Ca-WKY induced by NE, and the effect on the former was more significant than that on the latter（P〈0.05）. Ceaclusiea： TFH, Que and Isor might decrease the levels of [Ca^2＋], in VSMCs by blocking both voltage-dependent calcium channels （VDC） and receptoroperated calcium channels （ROC） in physiological or pathological state, which may be one of the important mechanisms of their hypotensive and protective effects on target organs in patients with hypertension.

Comparative quantitative analysis of fruit oil from Hippophae rhamnoides(seabuckthorn) by qNMR, FTIR and GC–MS

Objective: To develop a qNMR method for quantitative analysis of triacylglycerols in fruit oil of Hippophae rhamnoides(seabuckthorn, SBT) and analyze commercial samples of SBT oils using GC–MS and FTIR.Methods: SBT fruit oil(IPHRFH) was extracted with hexane and the triglyceride(TAG) was isolated by vacuum liquid chromatography. Six different branded SBT oils purchased from e-commerce suppliers(Amazon) and in-house prepared SBT oil was analyzed by qNMR and fatty acyl composition of TAGs determined by using NMR. In-house oil was also analysed by GC–MS and FTIR spectroscopy.Results: The qNMR results showed that the oil contained 80.3% of triacylglycerol(TAG). The SBT oil TAGs comprised of linolenate 6.6%, palmitoleate/oleate 65.4%, and total saturated fatty acyl chain including palmitate 28% as determined by qNMR. GC–MS analysis revealed that the major acyl functionalities present in the TAG were palmitoleic acid 36.5%, oleic acid 12.9%, palmitic acid 21.2%, and linoleic acid 18%. Of the six commercial samples analyzed, samples from only one supplier(SW) were fruit oil;All others were the seed oils or mix of fruit oil and seed oil. The labels for samples except for the SW did not indicate whether it was fruit oil or seed oil.Conclusion: The results suggest that SBT oil should be analyzed by combination of GC–MS, FTIR and qNMR for factual content of free fatty acid or TAGs, which are chemically different in nature and affect the quality of oil. GC–MS showed the content of omega free fatty acids after hydrolysis, while qNMR and FTIR showed the content of TAGs. The major acyl functionalities found in SBT fruit oil TAGs are palmitoleate/palmitate/oleate, while linoleate and linonelate make up a minor fraction. Furthermore,analysis of commercial samples showed discrepancies between label claims and actual content.

气相色谱内标法测定沙棘果肉油和种子油中的主要脂肪酸

以5个沙棘(Hippopha觕rhamnoides L.)品种的果实为试材,建立气相色谱内标法测定沙棘果肉油和种子油主要脂肪酸成分的方法。果肉油和种子油采用氯仿-甲醇法提取,甲醇钠-甲醇法甲酯化衍生后进行定性和定量。结果表明,十九烷酸甲酯是气相色谱法测定沙棘果实中脂肪酸成分含量的理想内标物。果肉油和种子油中的主要脂肪酸为棕榈酸、棕榈油酸、硬脂酸、油酸、亚油酸和α-亚麻酸。果肉油中棕榈油酸(36.25%)和棕榈油酸(28.92%)含量高,α-亚麻酸含量低;种子油中亚油酸(33.65%)和α-亚麻酸(32.95%)含量高,棕榈油酸含量低。5个品种沙棘的果肉油和种子油中主要脂肪酸的组成及含量存在一定的差异。亚历山大12号果肉油中的脂肪酸含量最高,为84.84 g/100 g油;无刺丰种子油中的脂肪酸含量最高,为88.39 g/100 g油。该方法操作简便、快速、重复性好,适合沙棘油脂样品中脂肪酸的定量测定。

沙棘良种‘红棘1号’快速繁育技术

‘红棘1号’沙棘(Hippophae rhamnoides L.)于2022年通过了国家林业和草原局的良种审定,被确定为林木良种用于林业生产。‘红棘1号’沙棘快速繁育技术,有利于沙棘产业的快速发展。本文从苗床建立、抽穗制备、扦插与苗床管理、移植等4个方面进行了系统阐述,为沙棘良种嫩枝扦插实现规范化和工厂化育苗提供技术支撑。

沙棘硬枝与嫩枝扦插扩繁技术

对大果沙棘(Hippophae rhamnoides L.)引进、杂交选育的沙棘优良品系的硬枝扦插和嫩枝扦插育苗技术进行了总结,分析了不同栽培区不同沙棘品系扦插繁殖的技术要点,发现硬枝扦插生根率在50%以上,嫩枝扦插生根率在不同试点连续多年均达75%以上;嫩枝扦插苗的移栽环节非常关键,要及时移栽;不同品种嫩枝生根能力的差异主要是品种的遗传特性和插穗的差异造成的;插穗的下部径的粗细与生根能力密切相关,建议采集的新梢插穗下部径应不小于0.3cm。

沙棘HrCYP85A1基因生物信息及表达分析

沙棘具有重要的生态价值及经济价值,油菜素内酯(BR)对植物抵御非生物胁迫具有响应作用,同时,CYP85A1是参与油菜素内酯合成反应的关键基因。因此,为了解析HrCYP85A1基因在沙棘抵御干旱胁迫中的作用,本研究以中国沙棘为试验材料,利用生物信息学软件对HrCYP85A1基因特点及蛋白结构进行了分析,并且利用荧光定量PCR技术分析了在干旱胁迫下HrCYP85A1基因的表达。结果表明,HrCYP85A1基因具有3个基序(motif),与多个物种的CYP85A1序列具有高度同源性;HrCYP85A1基因编码的蛋白位于质膜位置,且其为亲水性蛋白;HrCYP85A1参与NaCl、PEG与ABA等非生物胁迫的响应,说明其与沙棘多种非生物胁迫密切相关。本研究结果将为今后研究沙棘CYP家族基因功能提供了依据,并为林木抗逆研究及分子育种提供科学依据。