Production of polyhydroxybutyrate in switchgrass, a value-added co-product in an important lignocellulosic biomass crop

Polyhydroxyalkanoate bio-based plastics made from renewable resources can reduce petroleum consumption and decrease plastic waste disposal issues as they are inherently biodegradable in soil, compost and marine environments. In this paper, the successful engineering of the biomass crop switchgrass ( Panicum virgatum L.) for the synthesis of polyhydroxybutyrate (PHB) is reported. Polymer production was monitored in more than 400 primary transformants grown under in vitro and glasshouse conditions. Plants containing up to 3.72% dry weight of PHB in leaf tissues and 1.23% dry weight of PHB in whole tillers were obtained. Results from the analysis of the polymer distribution at the cellular and whole plant levels are presented, and target areas for the improvement of PHB production are highlighted. Polymer accumulation was also analysed in the T₁ generation obtained from controlled crosses of transgenic plants. This study presents the first successful expression of a functional multigene pathway in switchgrass, and demonstrates that this high-yielding biomass crop is amenable to the complex metabolic engineering strategies necessary to produce high-value biomaterials with lignocellulose-derived biofuels.     

Switchgrass nitrogen response and estimated production costs on diverse sites

Switchgrass (Panicum virgatum L.) has been the principal perennial herbaceous crop investigated for bioenergy production in North America given its high production potential, relatively low input requirements, and potential suitability for use on marginal lands. Few large trials have determined switchgrass yields at field scale on marginal lands, including analysis of production costs. Thus, a field‐scale study was conducted to develop realistic yield and cost estimates for diverse regions of the USA. Objectives included measuring switchgrass response to fertility treatments (0, 56, and 112 kg N ha^?1) and generating corresponding estimates of production costs for sites with diverse soil and climatic conditions. Trials occurred in Iowa, New York, Oklahoma, South Dakota, and Virginia, USA. Cultivars and management practices were site specific, and field‐scale equipment was used for all management practices. Input costs were estimated using final harvest‐year (2015) prices, and equipment operation costs were estimated with the MachData model ($2015). Switchgrass yields generally were below those reported elsewhere, averaging 6.3 Mg ha^?1 across sites and treatments. Establishment stand percent ranged from 28% to 76% and was linked to initial year production. No response to N was observed at any site in the first production year. In subsequent seasons, N generally increased yields on well‐drained soils; however, responses to N were nil or negative on less well‐drained soils. Greatest percent increases in response to 112 kg N ha^?1 were 57% and 76% on well‐drained South Dakota and Virginia sites, where breakeven prices to justify N applications were over $70 and $63 Mg^?1, respectively. For some sites, typically promoted N application rates may be economically unjustified; it remains unknown whether a bioenergy industry can support the breakeven prices estimated for sites where N inputs had positive effects on switchgrass yield.     

大黍(Panicum maximum Jacq.)的胚珠附器

观察了大黍(Panicum,maximum Jacq．)胚珠附器的发生时间、位置和发育过程及其细胞化学特征。结果显示：(1)大孢子母细胞时期,珠孔端有一个或多个珠心表皮细胞开始伸长、膨大,特化为胚珠附器。(2)当胚珠附器伸长、膨大至最大程度时,胚珠附器细胞表现出显著的极性特征：细胞核位于细胞的珠孔端,大而清晰;细胞内同时形成了一个特大的液泡,几乎占据了整个细胞的合点端;细胞质则被挤到珠孔端一侧,集中分布在核的周围。(3)胚珠附器从开始出现到发育成熟,都没有淀粉粒的积累;但是,PAS反应显示胚珠附器细胞壁和细胞质都比普通珠心细胞的染色程度深,这说明其细胞壁和细胞内部富含可溶性多糖。     

The influence of osmotic pretreatment and inoculum age on the initiation and regenerability of switchgrass suspension cultures

Summary Experiments were conducted to study the influence of osmotic pretreatment and inoculum (callus) age on the initiation and induction of somatic embryogenesis from suspension cultures of ‘Alamo’ switchgrass (Panicum virgatum L.). Embryogenic 10-, 20-, or 30-d-old calluses (hereafter referred to as inocula), produced from in vitro-cultured inflorescences, were used as explants to initiate the suspensions. Inocula were cultured for 30 h on MS solid medium containing 0.1, 0.2 or 0.3 M each of sorbitol and mannitol as an osmotic pretreatment. They were then transferred to liquid MS medium with 5 μM 6-benzylaminopurine and 9 μM 2,4-dichlorophenoxyacetic acid and cultured for 28 d in 2-ml Multiwell™ plantes. Individual multiwell contents were transferred to 125-ml Erlenmeyer flasks containing 20 ml of the above liquid medium and cultured for an additional 2 wk. Cultures initiated from the 10-d-old inoculum produced a higher embryogenic response than those initiated from 20- or 30-d-old inocula. Cultures initiated with 30-d-old inocula produced nonembryogenic clusters and the number of embryogenic cells was low. More embryos and a higher regeneration frequency were produced by 0.3 M than by 0.1 or 0.2 M of each of the osmostica.     

水培条件下pH值对柳枝稷幼苗生长发育的影响

土壤的p H值是限制植物生长发育的一个关键因素。柳枝稷是一种可作为牧草,水土保持的多年生C4能源植物。试验在水培条件下,利用裂区试验设计,以柳枝稷品种(系)为主区,水培液p H值为副区,对反映柳枝稷幼苗生长发育状况的指标进行统计分析。结果显示,随水培液p H值的酸碱强度增大,柳枝稷不同品种(系)幼苗的分蘖数、株高、苗鲜重、根冠比、根系活力以及净光合速率都极显著降低(P0.01),而幼苗保护酶活性以及丙二醛(MDA)含量则极显著升高(P0.01)。尤其是当PH值低于5.0时,幼苗的受到的胁迫更为明显,幼苗超氧化物歧化酶(SOD)和过氧化物酶(POD)活性反而急剧下降。与酸胁迫(p H值7.0)相比,柳枝稷对碱胁迫(p H值7.0)的适应能力更强,其中以西稷2号的抗逆性表现最好。因此应用柳枝稷在边际土壤地区推广种植时,要尽量避免强酸性土壤(p H值5.0),且选用西稷2号品系较为适宜。     

不同生境下铺地黍叶片蛋白质合成的比较

铺地黍是广泛分布在热带与亚热带的一种多年生禾草,可作为牧草及草坪植物,常见于农田、水湿地及海边,对盐度、干旱有较强的适应能力,是厦门岛滨海沙生植被的主要种类之一［１］。迄今尚未见对其耐盐（旱）机理的研究报道。本文通过比较铺地黍在不同生境下蛋白质组分的变化,从一个侧面探讨铺地黍的耐盐（旱）机理。１　材料和方法滨海沙生的铺地黍（ＰａｎｉｃｕｍｒｅｐｅｎｓＬ．）采自福建省厦门岛（东经１１７°５３′～１１８°２５′,北纬２４°２３′～２５°５５′）东海岸。该处沙滩地势平缓,坡度２０～２５度,盐度１．５％…     

外来植物坚尼草生态学研究

坚尼草（Panicm maximum Jacq.^-）是一种外来草本植物,已逸为野生,扩展迅速,对本地草本植物构成一定的影响。其扩展能力与其生长和繁殖特点有很大关系。研究表明,坚尼草对环境的适应能力较强;植株较高大,生长迅速,有较强的竞争能力;花期长,单丛坚尼草从始穗到终穗,花期可长达6－7个月;同一穗的果实熟期不同,而且边熟边脱落,从抽穗的5－7d起开始有果实脱落,脱落时间可持续1个月左右,以抽穗2－3周为脱落高峰;不同时间成熟的果实,明发率也不同,7月份成熟的果实比9成熟的果实萌发率高。同一穗中,以抽穗后2－3周（果实脱落的高峰期）成熟的果实萌发率较高。果实成熟后,不能立即萌发,储存1－6个月,种子萌发率较高。     

施氮和供水对混播和单播白羊草叶片叶绿素荧光特性的影响

为明确干旱条件下混播和施氮对白羊草〔Bothriochloa ischaemum(Linn.)Keng〕叶片叶绿素荧光参数的影响,采用盆栽法并设置不同混播比例〔白羊草与柳枝稷(Panicum virgatum Linn.)混播比例分别为8:0、6:2、4:4和2:6〕、施氮水平(即不施氮和1kg干土施01g纯氮)和供水条件(即正常供水和干旱胁迫6d后复水),对白羊草叶片叶绿素荧光参数的变化进行比较分析;在此基础上,采用一般线性模型分析这3个因素及其交互作用对白羊草叶绿素荧光参数的影响效应.结果显示:正常供水条件下,各处理组白羊草的最大光化学效率(Fv/Fm)、实际光化学效率(ΦPSⅡ)、表观光合电子传递速率(ETR)、光化学淬灭系数(qP)和非光化学淬灭系数(NPQ)均无明显变化.干旱及复水条件下,不施氮处理组白羊草的Fv/Fm值在干旱胁迫6 d(即土壤相对含水量最低)时降至最低值,并在复水2 d后恢复至与正常供水条件下相近的水平,而施氮处理组的Fv/Fm值则一直保持与正常供水条件下相近的水平;不论施氮与否,各处理组白羊草的ΦPSⅡ、ETR、qP和NPQ值基本上均表现为在干旱胁迫6 d时达到最高值,并在复水2 d后恢复至正常供水条件下各参数值的90%以上.总体上看,混播白羊草的qP和ETR值均高于单播白羊草,而其NPQ值则低于后者.统计分析结果表明:混播比例、施氮水平和供水条件3个因素间的交互作用对白羊草的ΦPSⅡ、qP和ETR值无显著影响,施氮水平对NPQ值的单独作用、施氮水平和混播比例的交互作用对Fv/Fm值以及施氮水平和供水条件的交互作用对ETR值也无显著影响,但这3个因素的单独作用及两两因素间的交互作用对白羊草其余叶绿素荧光参数均有显著或极显著影响.研究结果表明:一定程度的干旱胁迫有利于提高白羊草叶片PSⅡ反应中心的开放程度、光合电子传递速率和热耗散过剩光能的能力;在干旱胁迫条件下,施氮有助于白羊草叶片维持PSⅡ反应中心的活性和光化学效率;并且,与柳枝稷适度混播可改善白羊草叶片的光合性能,提高其种间竞争适应性.     

两种高质牧草不同生育期光合生理日变化及光响应特征

于2010年5月25日(拔节、分枝期)、7月6日(抽穗、结荚期)、8月25日(开花期)晴朗天气,采用LI-6400便携式光合作用测定系统对新引1号东方山羊豆和柳枝稷各项光合生理生态指标进行了测定。结果表明:(1)新引1号东方山羊豆净光合速率(Pn)日变化呈双峰曲线,存在"光合午休"现象;分枝期Pn、光能利用效率(LUE)、水分利用效率(WUE)日均值高于结荚期,而结荚期的叶温(Tl)、蒸腾速率(Tr)、气孔导度(Gs)、胞间CO2浓度(Ci)日均值高于分枝期。(2)柳枝稷的Pn日变化呈单峰曲线,拔节期Pn、Tr日均值高于抽穗期,开花期最低;WUE与Ci日均值在开花期最高,拔节期次之,抽穗期最小;Tl、LUE日均值在抽穗期最高,拔节期次之,开花期最小。(3)Pn与Tr、Gs、叶片气孔限制值(Ls)、光合有效辐射(PAR)具有极显著正相关关系,与Ci呈极显著负相关;Tr与Gs、叶温下蒸汽压亏缺(Vpdl)、气温(Ta)、Tl、PAR均呈极显著正相关关系,与大气CO2浓度(Ca)呈显著负相关;Tl、Gs与PAR呈极显著正相关。(4)通过光响应曲线的绘制及相关生理指标的计算,结合相关评价标准,得出柳枝稷为阳生植物,而新引1号东方山羊豆为耐荫植物,且柳枝稷比新引1号东方山羊豆对光环境的适应性强。     

半干旱黄土丘陵沟壑区柳枝稷（Panicum virgatum）的生物质形成

为深入认识半干旱黄土丘陵沟壑区引种能源植物柳枝稷生物质生产的开发潜力及其约束机制,调查了农田、植丛尺度上早熟和晚熟柳枝稷年度生命周期内生物量累积、分株建成动态,以及土壤水分供求平衡过程。研究发现,植丛尺度早熟柳枝稷抽穗比例近100％,分株生殖发生大小阈值依赖基本丧失,高度大小分布近似正态,种群内光资源竞争强度明显弱化,与晚熟类型形成明显差异。农田尺度晚熟柳枝稷生物质产量可以达到15t／hm^2,高出早熟类型近1倍,但是其立地80～400cm土层的含水量稳定在7％以下,土壤干旱已经发生,早期干旱胁迫导致的生长停滞,以及生长中后期的成片倒伏现象在两年的观测周期内连续出现。早熟柳枝稷立地则形成相对稳定的白草、柳枝稷复合优势植被结构,深层土壤水分含量稳定在10％以上,实现了跨年度的土壤水分供求平衡。植丛尺度的生物质形成在一定程度上取决于分株生殖发生的大小依赖程度和分株间竞争关系格局,基于植丛尺度普遍的生殖发生和明显弱化的光资源竞争,早熟柳枝稷表现出更为高效的生物质形成机制。农田尺度晚熟柳枝稷尽管在雨热同步期的降水资源利用效率上存在明显比较优势,但是在降水资源利用分配策略和效应上,早熟柳枝稷表现出了综合的生态适宜性优势。保证雨热同步期降水资源利用和保蓄的平衡,是半干旱黄土丘陵沟壑区生物质生产应该遵循的基本原则之一。