Germplasm Variability and Environmental Effects on Stem Cellulose and Lignin Concentrations in Alfalfa*

Stem cellulose and lignin concentrations are major determining factors of alfalfa (Medicago sativa ssp. sativa L.) forage quality. Only limited information is available on the genetic variability and the influence of environmental effects on these two stem-quality traits. Our objectives were to: evaluate the variability for stem cellulose and lignin concentrations in modern alfalfa germplasms across several harvests; observe the environmental stability of these two quality traits in 32 alfalfa clones selected high or low for either stem cellulose or lignin concentration; and examine the relationships between these two stem-quality traits and leaf and stem crude protein (CP). Fifty alfalfa entries (cultivars and experimental populations) were established May 1993, and sampled for stem acid-detergent lignin (ADL), stem acid-detergent cellulose (ADC), and stem CP on Sep 1993, June and Aug 1994. Clones were vegetatively propagated from individual plants selected for extremes in stem ADL and ADC and transplanted into blocks at two locations in May 1994 and sampled for quality analysis in September 1994. For all samples, leaves were hand-separated from stems and stem ADL, ADC, CP and leaf CP concentration were determined using near infrared reflectance spectroscopy. Entry differences for stem ADL and ADC were detected only at the June 1994 harvest date in the cultivar study. Spearman's ranked correlations over years in the clonal study demonstrated greater environmental stability for stem ADC (r = 0.70, P ≥ 0.01) than for stem ADL (r = 0.54, P # 0.05). Environmental effects had an impact on both traits, but stem ADC showed greater potential for improving forage quality. Simple correlations showed that decreasing stem ADL or ADC would have minimal effect on leaf CP and may increase stem CP.     

Effect of Leaf Sugar and Starch Concentrations on Apparent Photosynthesis in Alfalfa

Rates of apparent photosynthesis (AP), dark respiration, photorespiration, mesophyll resistance, and concentrations of soluble starch, sugars, chlorophyll and nitrogen were measured in alfalfa leaves ( Medicago sativa L.) grown under high or low light levels in an attempt to evaluate the effect of nonstructural carbohydrate concentration on net CO₂ assimilation.
Leaf N and chlorophyll levels of high light plants exceeded those of low-light plants. Constant light increased both starch and sugar levels in low-light plants, but had little effect on high-light plants. Rates of AP declined in low-light plants upon transfer to constant light conditions, apparently due to increases in stomatal and mesophyll resistance. Leaf starch levels were negatively correlated (P > 0.05) with rates of AP and positively correlated with mesophyll resistance in both groups of plants but leaf sugar content was not correlated with either AP or mesophyll resistance.
Starch and sugar levels of high-light plants declined substantially upon transfer to constant darkness, while low-light plants were less affected. Rates of AP declined in both groups of plants due to increases in both mesophyll and stomatal resistances. Neither sugar nor starch levels were correlated with AP or mesophyll conductance in this treatment.
The data support the hypothesis of feedback inhibition of photosynthesis, mediated through an increase in mesophyll resistance, which is associated with high levels of starch in the leaves.     

硒胁迫对紫花苜蓿硒积累及矿质元素吸收的效应

为了探讨硒胁迫对紫花苜蓿硒积累及对其他矿质元素吸收的影响。以3种苜蓿阿尔冈金、大叶苜蓿、维多利亚为试材,采用土培试验方法,研究了硒胁迫对紫花苜蓿不同组织的生物量、转运系数、耐性指数、硒积累量、硒含量和矿质元素含量的影响及紫花苜蓿对硒的积累能力。结果表明:施以100μmol/L硒时,硒处理显著提高了3个品种苜蓿根、茎、叶片的硒含量、耐性指数、硒积累量和K、P、Ca、Mg元素的吸收,当硒浓度为900μmol/L时,则降低了阿尔冈金和维多利亚不同器官生物量及大叶苜蓿根的生物量积累,且降低了阿尔冈金和维多利亚不同器官及大叶苜蓿根和茎对Zn吸收积累量。不同品种间相比较,大叶苜蓿生物量、硒积累量和K、P、Mg元素吸收显著高于其他2个苜蓿,且差异显著。综合评价表明,大叶苜蓿对矿质元素和Se有较好的吸收和富集能力。     

沙引发对紫花苜蓿种子盐逆境下发芽及幼苗生理生化变化的影响

试验以紫花苜蓿品种维多利亚和金皇后种子为材料，研究了沙引发处理对高盐(0．8％NaCl)逆境下种子发芽及幼苗生理生化变化的影响。结果表明，沙引发处理能显著提高紫花苜蓿两个品种种子的发芽率、发芽势、发芽指数和活力指数，缩短平均发芽时间，显著增加幼苗干重，同时提高了过氧化氢酶(CAT)、超氧化物歧化酶(SOD)和过氧化物酶(POD)活性，减少丙二醛(MDA)的积累。说明沙引发提高了紫花苜蓿种子的活力和抗盐胁迫能力，促进了盐逆境下种子的萌发和幼苗生长。     

不同品种紫花苜蓿中总皂苷含量的比较研究

采用比色法测定了北京地区栽培不同年限的45个紫花苜蓿品种茎、叶中总皂苷的含量。结果表明，各品种苜蓿叶片中的皂苷含量均高于茎，在不同品种之间存在明显差异。4年生植株中皂苷含量最高的品种“Vernal”为1.174%，最低的品种“Empress”为0.606%；3年生植株中最高的品种“WL442”为1.141%，最低的品种“农保”为0.788%；2年生植株中最高的品种“胜利者”为1.293%，最低的品种“全能”为0.722%。综合比较，含量最低品种与最高的品种相差2.13倍。栽培年限对紫花苜蓿皂苷含量具有一定影响。     

几种复混微肥对苜蓿微量元素营养的影响

根据河南省牧草种植区土壤微量元素含量、牧草营养及动物营养的需求特点,利用铁、锰、铜、锌、硼、钼、钴等几种微肥进行组配,来研究其对紫花苜蓿（Medicago sativa L.）微量元素营养的影响,其目的为生产提供不同用途的富含微量元素的优质牧草。结果显示：CD组配能促进紫花苜蓿第二茬铁、锌、硼,第三茬铁、锌、硼、铜含量的显著增加;及除第二茬钴外的各茬元素积累量增加。C1D1组配能促进紫花苜蓿第一茬钼、钴、硼,第二茬锌、钼、钴、硼、铁,第三茬铜、锌、硼、铁、锰含量的显著增加;及除第二茬铁外的各茬元素积累量增加。C1D2组配能促进紫花苜蓿第一茬锌、钼、钴,第二茬锌、钼、钴、硼、铁、锰,第三茬锌、钴、硼、铁、锰含量显著增加;及除第二茬铁外的各茬元素积累量的增加。C2D1组配能促进紫花苜蓿第一茬锌、钼、钴,第二茬铜、锌、钼、钴、硼、铁、锰,第三茬钼、硼、铁含量显著增加;及除第二茬铁外的各茬元素积累量的增加。C2D2组配能促进紫花苜蓿第一茬铜、锌、钼、钴,第二茬锰、铜、锌、钼、钴、硼、铁,第三茬锰、锌、钼、钴、硼、铁含量显著增加;及各茬元素积累量的增加。     

施肥对苜蓿现蕾期叶面积及比叶重的影响

用叶面积扫描仪测定苜蓿叶面积,在105℃下杀青,65℃下烘干24h,称取苜蓿叶干重,计算比叶重(SLW)。研究了不同施肥处理对3年生紫花苜蓿叶面积、叶干重及比叶重的影响。结果表明:①施肥对苜蓿现蕾期叶面积和叶干重都有一定的影响。K40P20配施不仅有利于扩大苜蓿光合叶面积,而且有利于苜蓿叶干重增加。②施肥,尤其是施氮提高了苜蓿现蕾期的比叶重,以K40P20N10配施效果最好,有助于苜蓿提高其光合速率,积累更多的光合产物。     

Primary Growth Parameters of Three Alfalfa Cultivars Adapted to Highland Climatic Conditions

An irrigated field study was conducted to determine the relative importance and inter‐relationships of growth parameters of three dormant alfalfa (Medicago sativa L.) cultivars grown in the highlands of Eastern Anatolia, Turkey, in 2000–2003. The fast‐growing cultivar Savas had the greatest dry matter (DM) yield at the final sampling date with the greatest mean crop growth rate. In addition to a greater relative growth rate (RGR), this cultivar had more stem branching and greater leaf area in the canopy, which resulted in greater leaf area index (LAI) and greater leaf area duration. Despite the considerable increase in leaf area ratio (LAR: leaf area per unit shoot DM), the reduction in RGR of all cultivars over time was the result of a large decline in net assimilation rate (NAR) due to increasing specific leaf area (SLA). Intracultivar variation in the RGR of alfalfa is mainly determined by NAR and SLA, and both were significantly higher for Savas than the other two cultivars. The relative importance of NAR and SLA to RGR changed due to increasing self‐shading as the LAI of the canopy increased, creating a trade‐off between NAR and SLA.     

德钦苜蓿品比试验报告

以阿尔冈金为对照品种,对德钦苜蓿进行了品种比较试验。结果表明：德钦苜蓿生育天数为233～246天。在3个试验点种植,越冬率平均为95.3%,与对照品种阿尔冈金（均值77.4%）差异显著（P<0.05）。德钦苜蓿和阿尔冈金三年3个试验点的平均株高分别为38.2 cm和27.1 cm,差异极显著（P<0.01）。其生长速度也与阿尔冈金差异极显著（P<0.01）。2004年三个试验点德钦苜蓿的茎叶比均显著低于阿尔冈金（P<0.05）,2005年、2006年均极显著低于阿尔冈金（P<0.01）。产量测定结果表明,三年各试验点德钦苜蓿鲜草、干草产量均极显著高于阿尔冈金（P<0.01）。营养成分分析表明,德钦苜蓿在初花期和盛花期粗干物质含量分别达25.73%和28.04%,蛋白含量分别达24.50%和19.41%,体外干物质消化率分别为68.03%和66.56%,其营养品质已经达到了1级水平。抗性分析表明,德钦苜蓿能较好地适应高温高湿环境,耐贫瘠。因而从引种的角度看,德钦苜蓿适应性强、高产、优质、适口性好,是一种值得在迪庆州推广种植的优良牧草。     

超量表达苹果酸脱氢酶基因提高苜蓿对铝毒的耐受性

在酸性土壤中,铝毒是许多作物正常生产的主要限制因子。虽可通过施用生石灰来改良土壤酸碱度,但只能改善土壤表层,在实际应用中受到很大限制。紫花苜蓿作为一种非常重要的牧草、饲料,在酸性土壤条件下苜蓿生长缓慢甚至死亡。我国南方水热资源丰富,但土壤多偏酸性,限制了苜蓿南移。有机酸能螯合铝离子,减轻铝对植物根系的危害。苹果酸是游离铝离子的有效螯合剂,而苹果酸脱氢酶(Malate Dehydrogenase,MDH)催化草酰乙酸形成苹果酸。本研究在苜蓿中超量表达苹果酸脱氢酶基因,通过抗生素筛选、组织化学染色和PCR扩增等技术鉴定出转化植株,并对转化株系进行了耐铝胁迫试验,比较分析了转基因株系与非转基因株系的的相对伸长量,转基因株系根相对伸长量比对照高出3．6％～22．5％。说明超量表达neMDH基因可提高了转基因苜蓿对铝毒的耐受性。     

水热胁迫对紫花苜蓿叶表皮蜡质组分及生理指标的影响

选用2个抗旱性不同的紫花苜蓿品种,敖汉(强抗旱)和三得利(弱抗旱),在水热胁迫条件下,调查其叶表皮蜡质含量及组分变化规律、蜡质含量与气体交换参数、脯氨酸及叶片相对含水量之间的关系。结果表明,紫花苜蓿叶表皮存在致密的蜡质层,蜡质晶体结构呈片状,无特殊的晶格方向。叶表皮蜡质主要由烷(1.98%~3.38%)、醇(79.97%~84.98%)、酯类(0.08%~0.24%)及其他少量未知物质组成(7.77%~13.38%)。品种类型、环境条件共同影响叶表皮蜡质的沉积。敖汉叶表皮蜡质含量显著高于三得利。水分胁迫后烷类比例增加(81.22%~108.16%),醇类比例下降(3.32%~12.54%),强抗旱品种叶表皮蜡质含量和气体交换参数无显著变化(除胞间二氧化碳浓度显著下降外),而弱抗旱品种蜡质含量和叶片光合速率、蒸腾速率、气孔导度及胞间二氧化碳浓度均显著下降。说明表皮蜡质限制水分散失,蜡质组分中烷类物质可能主要具限制水分散失的功能。高温及水热互作胁迫处理下,紫花苜蓿叶片光合速率和蒸腾速率下降,水分利用效率提高,叶片脯氨酸含量增加,相对含水量下降,敖汉蜡质含量下降,三得利蜡质含量无显著变化。表明在严重胁迫条件下紫花苜蓿主要通过关闭气孔和渗透调节来限制水分散失。     

Evaluation of R1 Alfalfa Somaclones for Herbage Yield and Morphological Characteristics

Tissue culture protocols were used to generate somaclonal variation in two synthetic populations of alfalfa. In both cases the somaclone populations give yields of dry matter about 29 % lower than the synthetic populations from which they were derived. Plant height and, for one population (LE-1), stem length, number of node per stem and internode length was also lower for the somaclones than for the original populations. The implications for plant breeding programs are considered and two hypotheses (one genetic and the other physiological), to account for the changes are presented.     

施肥对苜蓿开花期叶面积及比叶重的影响

【研究目的】研究了不同施肥处理对3年生紫花苜蓿开花期叶面积、叶干重及比叶重的影响。【方法】用叶面积扫描仪测定苜蓿叶面积,在105℃下杀青,65℃下烘干24h,称取苜蓿叶干重,计算比叶重(SLW)。【结果】①氮、磷、钾单施或不同方式的配施均能增加苜蓿开花期叶面积和叶干重。其中以K40P60配施最有利于扩大苜蓿光合叶面积。以K40P40N10配施最有利于提高苜蓿开花期叶干重。②施氮,尤其是氮与磷配施能明显提高苜蓿开花期的比叶重,以P20N10配施效果最好。而单施磷(P20、P40、P60)、单施钾(K20、K40)或K20P20配施均不利于增加苜蓿开花期比叶重。【结论】①苜蓿施K40P60最有利于扩大开花期光合叶面积,增强光合作用。②施K40P40N10最有利于提高苜蓿开花期叶干重,从而有利于晒制干草。③施P20N10能明显提高苜蓿开花期比叶重,有助于苜蓿提高其光合速率,积累更多的光合产物。     

锌锰微肥对苜蓿生产的效应研究

针对河南省气候土壤特点、锌锰钼肥的生理功能以及牧草生产利用中存在的问题,对Zn、Mn两种微肥在紫花苜蓿（Medicago sativa L.）优质牧草生产中的配施技术进行了研究。结果表明：紫花苜蓿的有效范围为400~500mg/kg,最佳浓度为430mg/kg。430mg/kg的Zn肥与380mg/kg的Mn 肥结合使用,效果更好,可使牧草产量提高约10.7%,种子产量提高13.7%。     

两个水稻叶色突变体的光合特性研究

以恢复系缙恢10号为对照,对它的两个叶色突变体ygl5和pygl1在孕穗期的光合、荧光特性和叶绿体超微结构进行了研究。结果表明,两个突变体叶绿体内基粒数量明显少于对照,叶绿素含量也大幅下降,其中ygl5表现为叶绿素总体缺乏,pygl1表现为叶绿素b严重缺乏;ygl5和pygl1的P_n、光饱和点、光补偿点、暗呼吸速率、表观量子效率和羧化效率都显著高于对照,而CO₂饱和点、CO₂补偿点和光呼吸速率则低于对照;其F_v/F_m、Φ_PSII和q_P均显著高于对照,表明突变体ygl5和pygl1具有较高的光能捕获效率和转换效率。     

绿肥作物箭筈豌豆种质产量性状综合评价

通过对124份绿肥箭筈豌豆种质资源品种产量性状指标进行主成分分析及聚类分析.结果表明,第3主成分(荚果数)偏高,第1主成分(株高)偏低,第2主成分(分枝数)适中,箭筈豌豆种子产量较高;第1,2主成分偏高,第3主成分偏低,即株高高,分枝数多的材料,干草产量高,可作为绿肥和饲用作物材料.聚类分析可综合评价箭筈豌豆的表现,发掘产量性状突出的材料,对于箭筈豌豆的育种具有重要的现实意义.     

紫花苜蓿锌指蛋白基因RNAi表达载体的构建及在苜蓿的转化

根据紫花苜蓿锌指蛋白MsZFN基因(GenBank登录号为EU624138)序列,设计两对含有酶切位点的特异性引物,以紫花苜蓿cDNA为模板,分别合成用于构建干扰载体的正反义片段,将正反义片段分别插入表达载体pART27的相应位置,构建成含有发夹结构的RNAi载体pART-F-R。利用农杆菌介导方法,将pART-F-R转化到紫花苜蓿中,经过PCR检测,获得了3株转基因植株。经过RT-PCR检测,证明转基因植株中MsZFN基因表达量明显低于未转基因的植株。结果表明,已构建成功具有发夹结构的RNAi载体pART-F-R,它可有效的抑制紫花苜蓿MsZFN基因。     

喷施硫酸钴对紫花苜蓿鲜草产量及品质的影响

采用叶面喷施的方法,研究了不同施钴水平对紫花苜蓿鲜草产量和营养品质的影响。结果表明：喷施硫酸钴显著（P＜0.05）提高了苜蓿鲜草产量,其中施钴量为300 mg/kg的处理效果最好,比对照CK产量可以提高25%。同时显著能够提高苜蓿叶茎比。不同施钴水平对苜蓿营养成分的含量影响不同,苜蓿粗蛋白及磷含量随随钴量的增加而增加,而对苜蓿粗纤维、粗灰分和无氮浸出物含量没有显著的影响,粗脂肪则随着施钴量的增加而逐渐降低。不同施钴水平对紫花苜蓿营养物质产量影响不同,其中粗蛋白、粗灰分、磷和无氮浸出物产量均以Co2处理300 mg/kg最高,都要显著高于对照,粗纤维、钙产量也以Co2处理最高,但与CK没有显著差异,粗脂肪产量则以Co1处理100 mg/kg效果最好     

Effect of Overseeding Forages into Alfalfa on Alfalfa Weevil, Forage Yield and Quality

Many alfalfa ( Medicago sativa L.) fields are sprayed each year with insecticides to control Egyptian alfalfa weevil ( Hypera brunneipennis ) and the alfalfa weevil ( Hypera postica ). Overseeding various species of legumes and grasses into established alfalfa has been proposed as a method to mitigate weevil damage without insecticides. Established fields of alfalfa were overseeded with legumes and grasses in randomized complete block designs in three years of study in a Mediterranean climate in the Sacramento Valley of California, USA. Several overseeding practices maintained or increased yields compared with the insecticide-sprayed treatment. Overseeding increased or maintained forage yield, eliminating the need for insecticides. However, overseeding did not prevent weevil damage to alfalfa. Overseeding is best used to extend the life of alfalfa stands in the final year of production, since overseeding can thin alfalfa stands. Market acceptance for mixed-species hay and harvest management of overseeded forages are the most important limitations to overseeding practices.     

Residual Effects of Rhizobium and Preplant N Fertilizer on Newly Established Alfalfa1

In areas where ineffective nodulation may be a problem, ‘starter’ nitrogen fertilization is often suggested as a useful management tool to sustain ineffectively nodulated seedling alfalfa (Medicago sativa L.) until an effective population of Rhizobium can develop. Although ineffective nodulation has been reported from several locations around the world, there have been no studies examing this possibility. A field experiment was conducted during 1982 and 1983 on a Woodburn silt loam soil (Aquultic Argixeroll) which contained a symbiotically ineffective native population of Rhizobium meliloti Dang. This study examined the residual and interactive effects of preplant N fertilizer (0, 45, 90 kg N ha^?1) and inoculation on the N nutrition and yield of seedling alfalfa during the first year of growth. The sustaining effects of the fertilizer N were transient and resulted in decreased N₂ fixation until 35 days following harvest. It was concluded that during the establishment year, managerial emphasis should be placed on inoculation and not on N fertilization to obtain a consistent, economical source of N for herbage protein and dry matter production.