海栖热袍菌耐高温β-甘露糖苷酶基因的克隆及表达

以海栖热袍菌(Thermotoga maritima)MSB8基因组DNA为模板,克隆得到β-甘露糖苷酶基因(man2).测序分析表明,该基因全序列为2358bp,编码785个氨基酸,分子量约为92×103.根据蛋白质氨基酸的同源性分析,该β-甘露糖苷酶与新阿波罗栖热袍菌(Thermotoga neapolitana)Man2(accession No.AAK52304.1)的同源性最大,达到80%.将此基因连接至表达载体pET-28a( )并转化到大肠杆菌BL21细胞中,经IPTG诱导,β-甘露糖苷酶活力达5.96U/mL.粗酶的温度稳定性分析表明,该酶的热稳定性好,90℃处理10min,活力回收率65%,具有重要的工业应用前景.图3表1参17     

木聚糖酶基因xynB在不同大肠杆菌表达系统中的表达比较

从黑曲霉(Aspergillus niger)nl-1中克隆获得木聚糖酶基因xynB.该基因全长745 bp,含有67 bp内含子,与公布的黑曲霉xynB基因有较高的同源性.将PCR扩增的木聚糖酶成熟肽基因和含有信号肽的基因分别连接到表达载体肠杆菌Top10 F’、DH10B和两种BL21宿主中获得重组菌株.通过IPTG诱导,xynB基因在重组菌株中获得特异性表达.表达产物以胞内可溶性蛋白和不溶性包涵体形式存在.诱导4 h,重组菌株pTrc-99a-xynB[BL21 condon plus(DE3)]表达量最高,胞内酶活达到299 IU/L.重组质粒pTrc-99a-xyn B(S)在不同宿主胞内和胞外均能分泌目的蛋白,诱导10 h,胞外酶活pTrc-99a-xynB(S)[BL21 condon plus(DE3)]达到347 IU/L.而pET-20b-xynB(S)在两种BL21宿主中均不表达.经SDS-PAGE分析,以pTrc-99a和pET-20b作为表达载体时,重组蛋白相对分子质量分别约为45×103和30×103.与pET-20b相比,pTrc-99a以及自身信号肽的引导更有利于重组木聚糖酶的表达.     

苏云金芽孢杆菌新基因cry1Ab17的克隆和生物信息学

利用高效克隆PCR产物的专用载体pMD18T,直接从苏云金芽孢杆菌WB9的PCR产物中克隆了cry1Ab17新基因.测序结果表明,该基因(GenBank登录号为AY646166)由3471个碱基组成,其编码的蛋白质含有1156个氨基酸残基,其中亲水性氨基酸占30.8%,疏水性氨基酸占45.2%,酸性氨基酸占12.9%,碱性氨基酸占11.1%.氨基酸序列的同源性分析结果表明,Cry1Ab17蛋白与已报道的Cry1Ab蛋白同源性为95.4%～99.7%,该蛋白的4个氨基酸残基———Pro170、Gly449、Gly796和Gly863与其它已报道Cry1Ab蛋白相应位置的氨基酸残基均不同.在核苷酸序列和氨基酸序列多重比较的基础上,应用PAUP4.0构建了Cry1A蛋白家族的系统发育树.SignalP分析结果显示,Cry1Ab17蛋白中不含信号肽序列.此外,对Cry1Ab17蛋白的二级结构和3个结构域也进行了预测和分析.图4表2参15     

解淀粉芽孢杆菌DC-4豆豉溶栓酶成熟肽编码序列的克隆及表达

利用PCR方法从解淀粉芽孢杆菌DC 4总DNA中扩增出豆豉溶栓酶 (DFE)成熟肽编码区片段 .测序结果表明 :DFE成熟肽编码区长 82 5bp,编码 2 75个氨基酸残基 ,分子量为 2 7.7× 10 3 ,推导的N’ -端氨基酸序列与豆豉溶栓酶N’ -端氨基酸测序结果完全一致 ,说明克隆到的基因确实是豆豉溶栓酶基因 .同源性分析表明 ,DFE成熟肽编码区的核苷酸和氨基酸序列与日本纳豆激酶的同源性分别为 80 .0 %和 86 .5 % ,这提示豆豉溶栓酶可能是一种新型的溶栓酶 .将表达质粒pET Nde转化E .coliBL2 1(DE3)中 ,IPTG可诱导表达大量的DFE融合蛋白 ,占菌体可溶性蛋白的 4 0 % ,主要以包涵体的形式存在 .图 3表 1参 19     

黑曲霉耐热木聚糖酶XYNB在毕赤酵母中的高效表达

高比活木聚糖酶的高效表达是进一步提高木聚糖酶发酵效价、降低生产成本的有效途径.将黑曲霉木聚糖酶基因XynB(不含信号肽)克隆到分泌型表达载体pPIC9K上,线性化后电击转化巴斯德毕赤酵母GS115,G418和PCR鉴定的阳性转化子经0.5%甲醇、在28℃诱导表达.SDS-PAGE分析表明,该蛋白相对分子质量为20×103左右.优化的诱导表达条件为,每隔12 h添加0.5%的甲醇,发酵5 d后,比活达4 757 U/mg;其最适温度为55℃,最适pH为5.0,80℃处理30min后仍有74%的残余酶活.     

簇毛麦(Dasypyrum villosum)GA20ox基因的分离和鉴定

通过RT-PCR技术克隆了簇毛麦(Dasypyrum villosum)赤霉素生物合成关键酶GA20ox基因的全长序列,命名为DvGA20ox,GenBank登录号为EU142949.该基因全长1 080 bp,推测编码359个氨基酸的蛋白质,具有植物GA20ox基因的典型保守结构域LPWKET和NYYPXCQKP.该基因推导编码蛋白质与小麦、大麦和黑麦草GA20ox蛋白的同源性分别为98%、97%和91%.该序列重组到原核表达载体pET-32a(+)获得的重组子pET-32a(+)-DvGA20ox转化大肠杆菌BL21pLysS后,IPTG诱导表达,SDS-PAGE分析表明,DvGA20ox基因在大肠杆菌中获得了高效表达,融合蛋白相对分子质量为55×103h,与理论值相符.结果为深入研究DvGA20ox蛋白的结构与功能以及该基因与株高发育的关系奠定了基础.图3参19     

麻疯树叶绿ω3脂肪酸脱饱和酶基因的克隆和鉴定

根据几个已知的植物ω3脂肪酸脱胞和酶基因序列,利用RT-PCR和RACE技术从麻疯树中克隆了一条编码叶绿体ω3脂肪酸脱饱和酶的cDNA序列.对该序列的分析表明,该cDNA的编码区长1 368bp,编码一个长度为455个氨基酸的蛋白质,推测的蛋白质相对分子质量(Mr)大小为52.1×103.同源分析显示,推测的氨基酸序列与其他物叶绿体脂肪酸脱饱和酶具有很高的相似性.RT-PCR分析表明,该基因在麻疯树叶片中有着稳定的表达.此外,还通过酵母表达系统鉴定了该基因编码的酶的功能.对转基因酵母的脂肪酸组成分析发现,α亚麻酸在转基因酵母中积累.上述结果表明,该麻疯树cDNA编码了一个叶绿体ω3脂肪酸脱饱和酶.     

香蕉几丁质酶基因ChiI2超表达载体和干涉表达载体的构建

分离克隆栽培香蕉中的几丁质酶基因ChiI2,构建超表达载体和干涉表达载体,可为进一步研究ChiI2基因响应抗逆胁迫的功能提供基础.从栽培香蕉天宝蕉(Musa spp., AAA)中克隆几丁质酶基因ChiI2,利用生物信息软件对该基因进行分析,并用无缝克隆技术分别构建ChiI2的超表达载体和干涉表达载体.从香蕉果皮中克隆到一个几丁质酶基因ChiI2,该基因全长942 bp,蛋白编码313个氨基酸,其编码的蛋白质理论分子量(Mr)为32.96,等电点(pI)为6.77.ChiI2蛋白的α-螺旋结构有6个,β-折叠结构有5个,转角结构有33个.蛋白质疏水性预测分析值为-0.233,属于亲水性蛋白.功能保守域分析表明,该蛋白是糖苷水解酶家族几丁质酶家族的一员.该基因核苷酸序列推导的氨基酸与野生香蕉、玉米、水稻、小麦、莲等植物的同源性都在70%以上.对ChiI2基因进行荧光定量PCR检测,发现4℃处理6 h的表达显著高于对照和38℃处理6 h,表明ChiI2基因可能与低温响应有关,诱导香蕉苗产生抗冷性.利用无缝克隆技术成功构建了pGreenII-ChiI2超表达载体和pGreenII-ChiI2i干涉表达载体,并转化到农杆菌EHA105菌株中.本研究成功地从栽培香蕉天宝蕉中分离克隆到了ChiI2基因,并对其基因特点和蛋白功能进行了预测分析,成功构建了超表达载体和干涉表达载体,可为进一步研究其功能奠定基础,也为采用基因工程方法改良选育香蕉抗寒品种提供了新尝试.(图8表1参24)     

甘蓝型油菜半胱氨酸蛋白酶cDNA的克隆及组织特异性表达分析

采用SSH (Suppression subtractive hybridization)和Race-PCR (Rapid-amplification of cDNA ends)技术克隆得到来自甘蓝型油菜(Brassica napus L.)矮化突变体'NDF-1'的一个差异表达基因编码区的全长cDNA序列,命名为Bncp5.该基因全长1 224 bp,含有1 080 bp的完整开放阅读框,编码359个氨基酸.该基因编码区与甘蓝中一个与衰老相关的半胱氨酸蛋白酶基因(登录号AF454960)同源性达97%,氨基酸序列同源性达到100%. Bncp5编码的蛋白相对分子质量为39.5×103,等电点为5.57.对其活性位点的分析表明其具备真核生物半胱氨酸蛋白酶的活性中心,且有很高的功能区段保守性,推测为一跨膜蛋白.相对定量PCR的检测结果表明, Bncp5在野生型和矮化突变体的根、茎、子叶和真叶中都有表达,但是表达量存在显著性差异.在野生型高秆油菜中,根的表达量最低,在真叶的表达量最高;在矮化突变体中,根、茎中的表达明显高于野生型,而在子叶和真叶中的表达量与野生型油菜的表达量基本一致.     

桃ACC合酶基因组DNA（pACSG01）的序列分析及其表达

以桃基因组DNA为模板,用套式PCR技术扩增并克隆了桃ACC合酶基因片段,将其克隆（定名为pACSG01）并进行序列测定,表明该自然全长1320bp,并富含HindⅢ和EcoRⅠ位点,与其它ACC合酶基因结构序列有一定的相似性,内含有两个内含子,编码的氨基酸序列与已克隆桃ACC合酶cDNA推导的氨基酸序列的同源性分别为57．4％和56．8％,pACSG01与我们已克隆的两个桃ACC合酶cDNA基因表达有所不同,RNA点杂交和RT－PCR结合Southern杂交分析表明,该基因在成熟和乙烯处理的果实均不表达,伤处理、LiCl和生长素处理也不能诱导核基因的表达,在衰老花瓣中也不表达,图3参18。     

Achieving Open Access to Conservation Science

Conservation science is a crisis discipline in which the results of scientific enquiry must be made available quickly to those implementing management. We assessed the extent to which scientific research published since the year 2000 in 20 conservation science journals is publicly available. Of the 19,207 papers published, 1,667 (8.68%) are freely downloadable from an official repository. Moreover, only 938 papers (4.88%) meet the standard definition of open access in which material can be freely reused providing attribution to the authors is given. This compares poorly with a comparable set of 20 evolutionary biology journals, where 31.93% of papers are freely downloadable and 7.49% are open access. Seventeen of the 20 conservation journals offer an open access option, but fewer than 5% of the papers are available through open access. The cost of accessing the full body of conservation science runs into tens of thousands of dollars per year for institutional subscribers, and many conservation practitioners cannot access pay‐per‐view science through their workplace. However, important initiatives such as Research4Life are making science available to organizations in developing countries. We urge authors of conservation science to pay for open access on a per‐article basis or to choose publication in open access journals, taking care to ensure the license allows reuse for any purpose providing attribution is given. Currently, it would cost $51 million to make all conservation science published since 2000 freely available by paying the open access fees currently levied to authors. Publishers of conservation journals might consider more cost effective models for open access and conservation‐oriented organizations running journals could consider a broader range of options for open access to nonmembers such as sponsorship of open access via membership fees. Obtención de Acceso Abierto a la Ciencia de la Conservación     

Some polyphagous Homoptera gain protection from their natural enemies by feeding on the toxic plantsSpartium junceum andErythrina corallodendrum (Leguminosae)

Summary Interactions were studied among alkaloid-containing legumes (Erythrina corallodendrum andSpartium junceum) and non-toxic plants (Citrus sinensis, Cucurbita moschata andEuphorbia tirucalli), several polyphagous homopterans,Aphis craccivora (Aphididae),Icerya purchasi, I. aegyptiaca (Margarodidae),Lepidosaphes ulmi (Diaspididae) andPlanococcus citri (Pseudococcidae), and some major natural enemies of these homopterans. Significant reductions in survival due to negative effects of alkaloid containing as compared with non-alkaloidal plants were recorded for the predatorsRodolia cardinalis andChilocorus bipustulatus, but not forCryptolaemus montrouzieri (Coleoptera: Coccinellidae),Chrysoperla carnea (Neuroptera: Chrysopidae) andSympherobius sanctus (Neuroptera: Sympherobiidae). The development time of the larvae or pupae ofR. cardinalis, C. carnea andS. sanctus was longer on the toxic plants than on the non-toxic ones. The percentage of parasitism ofA. craccivora collected from the non-alkaloidal plantsVicia palaestina andMelilotus albus was much higher than that onS. junceum. The parasitoid complexes ofA. craccivora differed between both plant groups. The nutritive value of honeydew ofI. purchasi andA. craccivora, as expressed by the life span ofEncyrtus infelix (Hymenoptera: Encyrtidae) adults, was also investigated. Life spans were significantly longer when the wasps fed on honeydew produced on non-alkaloidal plants (C. sinensis andPittosporum tobira) than on alkaloid containing plants whenI. purchasi — but notA. craccivora — was the producer. It is suggested that the chemical defense ofE. corallodendrum andS. junceum is exploited by polyphagous phytophages to reduce predation. In nature, population growth and density of four of the investigated homopterans are conspicuously high when they developed on the alkaloid containing plant species, and very low on non-alkaloid plants. The efficiency of their natural enemies may be reduced by sequestration of alkaloids (or other toxic plant compounds) or their transfer into excreted honeydew. Therefore it is assumed that a generalist phytophagous homopteran may be protected from its natural enemies, although at different rates of efficiency, if it can safely sequester the host allelochemical when it develops on toxic species within its host range.     

Rapid,recurring, structured survey versus bioblitz for generating biodiversity data and analysis with a multispecies abundance model

A bioblitz inexpensively and quickly generates biodiversity data, but bioblitzes are often conducted with haphazard, unreplicated sampling. Results tend to be taxonomically, geographically, or temporally biased, lack metadata, and consist of lists of observed taxa that do not enable further analyses or correction for imperfect detection. A rapid, recurring, structured survey (RRSS) uses a structured sampling design and temporal and spatial replication to survey randomly selected sites on a conservation property. We participated in a loosely structured bioblitz and a subsequent RRSS at Big Canoe Creek Nature Preserve in Springville (St. Clair County), Alabama (USA) to compare observed richness derived from the 2 survey approaches. The RRSS data structure enabled us to fit models that accounted for imperfect detection to estimate abundances, occupancy probabilities, and habitat associations. The loosely structured bioblitz data could not be used in such models. We present a new integrated multispecies abundance model that we applied to avian RRSS data. Our model extension enables estimation for the community, employs data augmentation to estimate the number of undetected species, and incorporates covariates. The RRSS generated a more comprehensive and less biased list of observed taxonomic richness than the loosely structured bioblitz (e.g., 73 vs. 45 bird species and 104 vs. 63 insect families from the RRSS vs. loosely structured bioblitz, respectively). Models fit to the RRSS data identified seasonal patterns in avian community composition and allowed for estimation of habitat–occupancy relationships for insect taxa. The RRSS protocol has potential for broad transferability as a standardized, quick, and inexpensive way to inventory biodiversity and estimate ecological parameters while providing an outreach opportunity.     

Sequestration of cucurbitacin analogs by New and Old World chrysomelid leaf beetles in the tribe Luperini

Summary Two South American polyphagous leaf beetles,Diabrotica speciosa andCerotoma arcuata, selectively accumulated the bitter tasting compound 23,24-dihydrocucurbitacin D in their body after ingesting root tissues of cucurbit plants. Similarly, three Asian Cucurbitaceae-feeding specialists in the genusAulacophora were found to sequester the same compound. Cucurbitacin analogs were shown to deter feeding by a bird predator, indicating an allomonal role for these compounds in cucurbitacin-associated chrysomelid leaf beetles both of New and Old Worlds. The strong affinity to cucurbitacins, selective sequestration of the analogs and consequent protection from predators suggested an ecological adaptation mechanism developed in common among these two geographically isolated subtribes in the Luperini.     

Effects of Controlled Fire and Livestock Grazing on Bird Communities in East African Savannas

Abstract: In East Africa fire and grazing by wild and domestic ungulates maintain savannas, and pastoralists historically set fires and herded livestock through the use of temporary corrals called bomas. In recent decades traditional pastoral practices have declined, and this may be affecting biodiversity. We investigated the effects of prescribed fires and bomas on savanna bird communities in East Africa during the first and second dry seasons of the year (respectively before and after the rains that mark the onset of breeding for most birds). We compared abundance, richness, and community composition on 9‐ha burned plots, recently abandoned bomas, and control plots in the undisturbed matrix habitat over a 3‐year period. Generally, recently burned areas and abandoned bomas attracted greater densities of birds and had different community assemblages than the surrounding matrix. The effects of disturbances were influenced by interactions between primary productivity, represented by the normalized difference vegetation index, and time. Bird densities were highest and a greater proportion of species was observed on burned plots in the months following the fires. Drought conditions equalized bird densities across treatments within 1 year, and individuals from a greater proportion of species were more commonly observed on abandoned bomas. Yearly fluctuations in abundance were less pronounced on bomas than on burns, which indicate that although fire may benefit birds in the short term, bomas may have a more‐lasting positive effect and provide resources during droughts. Several Palearctic migrants were attracted to burned plots regardless of rainfall, which indicates continued fire suppression may threaten their already‐declining populations. Most notably, the paucity of birds observed on the controls suggests that the current structure of the matrix developed as a result of fire suppression. Traditional pastoralism appears critical to the maintenance of avian diversity in these savannas.     

Compliance with and ecosystem function of biodiversity offsets in North American and European freshwaters

Land-use change via human development is a major driver of biodiversity loss. To reduce these impacts, billions of dollars are spent on biodiversity offsets. However, studies evaluating offset project effectiveness that examine components such as the overall compliance and function of projects remain rare. We reviewed 577 offsetting projects in freshwater ecosystems that included the metrics project size, type of aquatic system (e.g., wetland and creek), offsetting measure (e.g., enhancement, restoration, and creation), and an assessment of the projects’ compliance and functional success. Project information was obtained from scientific and government databases and gray literature. Despite considerable investment in offsetting projects, crucial problems persisted. Although compliance and function were related to each other, a high level of compliance did not guarantee a high degree of function. However, large projects relative to area had better function than small projects. Function improved when projects targeted productivity or specific ecosystem features and when multiple complementary management targets were in place. Restorative measures were more likely to achieve targets than creating entirely new ecosystems. Altogether the relationships we found highlight specific ecological processes that may help improve offsetting outcomes.     

A palaeo-hydrogeological model for arsenic contamination in southern and south-east Asia

An argument is presented in which areas of natural arsenic contamination of modern groundwaters throughout Asia have a common origin. Arsenic originally accumulated in oceanic ferro-manganoan sediments of the eastern Palaeo-Tethys. This was further concentrated through oceanic crustal extinction in what later became the south-east Chinese accreted mineralised terrain. Proto-Himalayan uplift of this area created the palaeo-drainage systems of the Ganges – Brahmaputra, Irrawaddy, Mekong, and Red Rivers, with consequent headwater erosion of arsenic-rich sediments. Their downstream deposition as immature and easily redistributed Neogene sandstones, silts, and iron-rich clays has created secondary and tertiary reservoirs of adsorbed and authigenic arsenic, from which the current arsenic-rich groundwaters have evolved. Considering river basins within the above palaeo-hydrogeological framework provides a basis for assessing the risk of arsenic in groundwater basins of south and south-eastern Asia.     

What Every Conservation Biologist Should Know about Economic Theory

Abstract: The last century has seen the ascendance of a core economic model, which we will refer to as Walrasian economics. This model is driven by the psychological assumptions that humans act only in a self‐referential and narrowly rational way and that production can be described as a self‐contained circular flow between firms and households. These assumptions have critical implications for the way economics is used to inform conservation biology. Yet the Walrasian model is inconsistent with a large body of empirical evidence about actual human behavior, and it violates a number of basic physical laws. Research in behavioral science and neuroscience shows that humans are uniquely social animals and not self‐centered rational economic beings. Economic production is subject to physical laws including the laws of thermodynamics and mass balance. In addition, some contemporary economic theory, spurred by exciting new research in human behavior and a wealth of data about the negative global impact of the human economy on natural systems, is moving toward a world view that places consumption and production squarely in its behavioral and biophysical context. We argue that abandoning the straightjacket of the Walrasian core is essential to further progress in understanding the complex, coupled interactions between the human economy and the natural world. We call for a new framework for economic theory and policy that is consistent with observed human behavior, recognizes the complex and frequently irreversible interaction between human and natural systems, and directly confronts the cumulative negative effects of the human economy on the Earth's life support systems. Biophysical economics and ecological economics are two emerging economic frameworks in this movement.     

Effects of Coffee Management on Deforestation Rates and Forest Integrity

Knowledge about how forest margins are utilized can be crucial for a general understanding of changes in forest cover, forest structure, and biodiversity across landscapes. We studied forest‐agriculture transitions in southwestern Ethiopia and hypothesized that the presence of coffee (Coffea arabica)decreases deforestation rates because of coffee's importance to local economies and its widespread occurrence in forests and forest margins. Using satellite images and elevation data, we compared changes in forest cover over 37 years (1973–2010) across elevations in 2 forest‐agriculture mosaic landscapes (1100 km² around Bonga and 3000 km² in Goma‐Gera). In the field in the Bonga area, we determined coffee cover and forest structure in 40 forest margins that differed in time since deforestation. Both the absolute and relative deforestation rates were lower at coffee‐growing elevations compared with at higher elevations (?10/20% vs. ?40/50% comparing relative rates at 1800 m asl and 2300–2500 m asl, respectively). Within the coffee‐growing elevation, the proportion of sites with high coffee cover (>20%) was significantly higher in stable margins (42% of sites that had been in the same location for the entire period) than in recently changed margins (0% of sites where expansion of annual crops had changed the margin). Disturbance level and forest structure did not differ between sites with 30% or 3% coffee. However, a growing body of literature on gradients of coffee management in Ethiopia reports coffee's negative effects on abundances of forest‐specialist species. Even if the presence of coffee slows down the conversion of forest to annual‐crop agriculture, there is a risk that an intensification of coffee management will still threaten forest biodiversity, including the genetic diversity of wild coffee. Conservation policy for Ethiopian forests thus needs to develop strategies that acknowledge that forests without coffee production may have higher deforestation risks than forests with coffee production and that forests with coffee production often have lower biodiversity value. Efectos de la Administración Cafetalera sobre las Tasas de Deforestación y la Integridad de los Bosques     

Threats to intact tropical peatlands and opportunities for their conservation

Large, intact areas of tropical peatland are highly threatened at a global scale by the expansion of commercial agriculture and other forms of economic development. Conserving peatlands on a landscape scale, with their hydrology intact, is of international conservation importance to preserve their distinctive biodiversity and ecosystem services and maintain their resilience to future environmental change. We explored threats to and opportunities for conserving remaining intact tropical peatlands; thus, we excluded peatlands of Indonesia and Malaysia, where extensive deforestation, drainage, and conversion to plantations means conservation in this region can protect only small fragments of the original ecosystem. We focused on a case study, the Pastaza‐Marañón Foreland Basin (PMFB) in Peru, which is among the largest known intact tropical peatland landscapes in the world and is representative of peatland vulnerability. Maintenance of the hydrological conditions critical for carbon storage and ecosystem function of peatlands is, in the PMFB, primarily threatened by expansion of commercial agriculture linked to new transport infrastructure that is facilitating access to remote areas. There remain opportunities in the PMFB and elsewhere to develop alternative, more sustainable land‐use practices. Although some of the peatlands in the PMFB fall within existing legally protected areas, this protection does not include the most carbon‐dense (domed pole forest) areas. New carbon‐based conservation instruments (e.g., REDD+, Green Climate Fund), developing markets for sustainable peatland products, transferring land title to local communities, and expanding protected areas offer pathways to increased protection for intact tropical peatlands in Amazonia and elsewhere, such as those in New Guinea and Central Africa which remain, for the moment, broadly beyond the frontier of commercial development.