DNA Barcoding Wild Flora in Pakistan’s Forests

DNA Barcoding Wild Flora in Pakistan’s Forests

DNA Barcoding Wild Flora in Pakistan’s Forests

Preserving voucher specimens and creating a virtual herbarium to understand and protect some of the oldest living trees on the planet.
Juniper Forest of Ziarat, Balochistan, Pakistan.
PHOTO CREDIT: Nazeer Ahmed
Balochistan, the largest province of Pakistan, is endowed with a variety of natural forests. Juniper (Juniperus excelsa), Pinus (Pinus gerardiana), wild Olive (Olea sp.) and mangroves are the predominant forest ecosystems of the province. The versatility of life forms in these forests support dynamic ecosystems and provide several important ‘ecosystem services’ like food, medicines, climate regulation, genetic resources, recreation facilities, etc. Biodiversity conservation, in the face of such benefits, becomes imperative. Comprehensive cataloguing of flora and fauna is, by all means, at the heart of such conservation endeavours.

The Juniper forest of Ziarat, Balochistan, declared a UNESCO Man and Biosphere Reserve, is considered one of the world’s largest compact forests of its kind spreading over an area of 100,000 ha. Being some of the oldest living trees on earth, they are termed “Living Fossils”. The Chilgoza (Pinus gerardiana), also known as the Chilgoza Pine, on the other hand, are listed as lower risk, near threatened forest. Anthropogenic interferences have further aggravated the situation in this ecosystem and a more focused study about their current status is needed.

Juniper Forest of Ziarat, Balochistan, Pakistan.
PHOTO CREDIT: Nazeer Ahmed

Fragmented studies exist attempting to document the associated flora of these forests; however, a more comprehensive approach is needed. The use of DNA barcoding techniques, duly augmented by classical taxonomy, is necessary for the creation of a reference library to inventory, assess, and describe the biodiversity of these forests. To fill this gap, a study was designed to provide a foundation for future biodiversity assessment and conservation efforts.

 

Funded by Pakistan Agricultural Research Council and Higher Education Commission of Pakistan, our research group at the Balochistan University of Information Technology, Engineering & Management Sciences, Quetta is expecting to barcode and acquire samples of approximately 1,000 wild plant species. 

To date, 730 samples of 525 different species have been collected and 29% (150 of 525) have been barcoded. Besides maintaining voucher specimens, a virtual herbarium will be made available to the global scientific community interested in the flora of these forest ecosystems.

Read more about Pakistan:

SMALL STEPS LEAD TO BIG INITIATIVES: PAKISTAN REAFFIRMS SUPPORT FOR IBOL BY LAUNCHING PAKBOL

From economically important insect species to plants to food security, Pakistani researchers are working to barcode all life in their country through a national initiative – PakBOL.

UNIVERSITY OF SINDH JAMSHORO BARCODES GRASSHOPPERS IN PAKISTAN’S THAR DESERT

Tracking the shift of non-pests to crop pests, a phenomenon accelerated by anthropogenic pressures in the Thar Desert.

Written by

Nazeer Ahmed

Nazeer Ahmed

Balochistan University of Information Technology, Quetta, Pakistan

April 7, 2019
PDF
https://doi.org/10.21083/ibol.v9i1.5476

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Scat Raiders Unravel Animal-Plant Interactions in Lebanon Using DNA Barcoding Tools

Scat Raiders Unravel Animal-Plant Interactions in Lebanon Using DNA Barcoding Tools

Scat Raiders Unravel Animal-Plant Interactions in Lebanon Using DNA Barcoding Tools

Using DNA dietary analysis on Eastern Mediterranean wildlife to explore the role of animals in ecological restoration processes.
Plant collection in Ehden Nature Reserve – north Lebanon. PHOTO CREDIT: Saint Joseph University

Lebanon is considered a hotspot for biodiversity in the Mediterranean basin likely due to its geographic position at the transition of two major landmasses (that is Eurasia and Africa). The Lebanese territory is divided between mountainous slopes with fertile valleys separating the two mountain chains that run parallel with the sea and the steppe areas in the north-east. Deep canyons and numerous rivers characterize this mountainous landscape.

These geomorphological regions give rise to many bio-climatic zones and several habitat types that are home to more than 9,116 described species (4,486 for fauna and 4,630 for flora from which 91 are endemic). However, major taxonomic groups like insects and fungi are understudied and taxa are underrepresented within public data platforms. For example, according to the Barcode of Life Data System (BOLD), only 345 Lebanese specimens with sequences are published, forming 151 BINs and, of these records, only 108 have species names.

In September 2018, the Faculty of Science at Saint Joseph University of Beirut joined the iBOL Consortium providing us with the opportunity to unravel Lebanese biodiversity by DNA barcoding both small and large mammals as well as the main trees and shrubs used in reforestation programs. We will also target endemic plant species.

Animals are a crucial component for the resilience of forest ecosystems and an important factor in forest restoration projects as they promote the sustainability of reintroduced plants, as well as seed dispersal. However, we still need to identify the animals present in restored areas.

Animal scat collection. PHOTO CREDIT: Saint Joseph University

In addition, knowing what each animal eats and which plant seeds are being dispersed is crucial for reforestation schemes that promote wildlife and ensure ecosystem sustainability. The information needed to study the diets of animals can be found hidden in their scat which contains not only the animal’s DNA, but also what that animal has eaten. With the powerful technique of DNA metabarcoding, we now have the necessary tool to efficiently unravel the genetic information hidden in animal scat. The DNA sequences obtained from such material are identified by comparison to a reference library of animals and plants of the Eastern Mediterranean countries.

 

Constructing the Reference Library – DNA isolation. PHOTO CREDIT: Saint Joseph University
This reference library was prepared from leaves collected in the wild and from DNA isolated from dead animals found along roads or from private museums. Thus, we have generated sequences for 51 plants and 18 mammals. This study conducted in collaboration with the Smithsonian Conservation Biology Institute and the University of Otago is the first to employ a DNA dietary analysis on wildlife in the Eastern Mediterranean Region and explicitly considering the role of wildlife in ecological restoration processes. Our results will inform management strategies to help with the conservation efforts of these imperiled species.

Written by

Carole Saliba

Carole Saliba

Faculty of Science, Saint-Joseph University

Liliane Boukhdoud

Liliane Boukhdoud

Faculty of Science, Saint-Joseph University

Magda Bou Dagher Kharrat

Magda Bou Dagher Kharrat

Faculty of Science, Saint-Joseph University

April 7, 2019
PDF
https://doi.org/10.21083/ibol.v9i1.5489

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HOW BIOSCAN IS INSPIRING THE NEXT GENERATION OF RESEARCHERS

They were enlightened by the idea of discovering new species and by the possibility of doing so using DNA barcoding tools.”

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DNA Barcoding and Genomics in the Megadiverse Amazon Altitude Fields

DNA Barcoding and Genomics in the Megadiverse Amazon Altitude Fields

DNA BARCODING AND GENOMICS IN THE MEGADIVERSE AMAZON ALTITUDE FIELDS

Scientists are contributing to the most profound molecular representation of biodiversity in any Brazilian environment.

The entrance of a ferruginous cave at the Bocaina mountain in the Carajás region, Pará State, Brazil.

PHOTO CREDIT: João Marcos Rosa.

Industrial activities in the Brazilian Amazon are highly regulated by governmental agencies. However, the lack of knowledge about megadiverse areas is a problem for the establishment of best conservation practices; this is the case for mining operations in the eastern Amazonian Carajás, a region comprised of a mosaic of national parks, indigenous peoples’ conservation areas, and nature reserves. Of particular interest are the ferruginous altitude fields known as the Canga. Our floristic survey described the presence of 1,094 species from just over 200 previously known1. The lack of biodiversity data is even more significant for the ferruginous caves where only around 10 invertebrate species are identified to the species level. Unfortunately, this is typical for the Amazon basin.

To provide reliable scientific data that contributes to the implementation of best conservation practices, Instituto Tecnológico Vale is developing DNA barcode reference libraries for the flora, cave invertebrates, and bats of the region, and providing deeper genomic references for species that are endangered or difficult to identify. To achieve this goal, we established the necessary infrastructure to conduct DNA sequencing using Sanger, Illumina, and PacBio technologies, coupled with high-performance computing, artificial intelligence algorithms, and highly trained personnel.

 

Botanists collecting samples in a temporary lake in ferruginous altitude fields in the Carajás region, Pará State, Brazil.
PHOTO CREDIT: João Marcos Rosa

To date, over 8,575 barcodes for 3,548 specimens of plants and invertebrates have been produced, while a large number of species remain to be identified by morphological attributes. Morphological specimen determination is conducted by in-house specialists, as well as by an extensive network of specialists in universities and museums across Brazil and abroad.

For this purpose, nuclear and chloroplast or mitochondrial markers as well as low coverage to whole genome sequencing or restriction site-associated DNA sequencing (RADSeq) are being employed to unravel the vast genetic diversity of the biota of Carajás2-4. For several endemic plants, such as species of Asteraceae, Melastomataceae5, Convolvulaceae6, and Isoetaceae2, diversity analyses, based on next-generation sequencing, aim to characterize the genetic variability among and within populations, as well as the identification of markers under selective pressure. These methods also contribute to the understanding of population structure and the process of gene flow between populations affected by natural factors and industrial operations. Models of environmental distribution, including parameters sensitive to climate change, were determined for several taxonomic groups, including plants and bats7.

Specimens of the pinheirinho-da-canga (Paepalanthus fasciculoides) highly adapted to inhabit the canga (ancient ferruginous rock outcrops) at the altitude fields in the Carajás region, Pará State, Brazil.
PHOTO CREDIT: João Marcos Rosa
We are also establishing eDNA methods, as well as metagenomics and metaproteomics data for environmental monitoring of ferruginous fields phytophysiognomies, areas under rehabilitation processes, and caves8. Together these data constitute the most profound molecular representation of any environment in Brazil. We have contributed a total of 3,072 specimens to the Barcode of Life Data System (BOLD) comprising 398 genera (291 new) in addition to the 408 different genera collected through the national effort for angiosperms in Brazil. We have also provided 571 cave fauna specimens.

It is important to highlight that all of these data generated are being provided to the public and its use will be critical to the conservation of such a unique collection of species.

References:

1. Brazil Flora Group (2018) Growing knowledge: an overview of Seed Plant diversity in Brazil. Rodriguésia 66(4): 1085–1113. http://dx.doi.org/10.1590/2175-7860201566411

2. Nunes GL, Oliveira RRM, Guimarães JTF, Giulietti AM, Caldeira C, Vasconcelos S, et al., (2018) Quillworts from the Amazon: A multidisciplinary populational study on Isoetes serracarajensis and Isoetes cangae. PLoS ONE 13(8): e0201417. https://doi.org/10.1371/journal.pone.0201417

3. Ramalho AJ, Zappi DC, Nunes GL, Watanabe MTC, Vasconcelos S, Dias MC, Jaffé R, Prous X, Giannini TC, Oliveira G and Giulietti AM (2018) Blind testing: DNA barcoding sheds light upon the identity of plant fragments as a subsidy for cave conservation. Frontiers in Plant Science 9:1052. https://doi.org/10.3389/fpls.2018.01052

4. Oliveira RRMO, Vasconcelos S, Pires ES, Pietrobon T, Prous X and Oliveira G (2019) Complete mitochondrial genomes of three troglophile cave spiders (Mesabolivar, pholcidae), Mitochondrial DNA Part B 4(1): 251–252. https://doi.org/10.1080/23802359.2018.1547139

5. Carvalho CdS, Lanes ECM, Silva AR, Caldeira CF, Carvalho-Filho N, Gastauer M, Imperatriz-Fonseca VL, Nascimento W, Oliveira G, Siqueira JO, Viana PL, Jaffe R (2019) Habitat loss does not always entail negative genetic consequences. bioRxiv 528430. https://doi.org/10.1101/528430

6. Lanes ÉC, Pope NS, Alves R, Carvalho Filho NM, Giannini TC, Giulietti AM, Imperatriz-Fonseca VL, Monteiro W, Oliveira G, Silva AR, Siqueira JO, Souza-Filho PW, Vasconcelos S and Jaffé R (2018) Landscape genomic conservation assessment of a narrow-endemic and a widespread morning glory from Amazonian Savannas. Frontiers in Plant Science 9:532. https://doi.org/10.3389/fpls.2018.00532

7. Costa WF, Ribeiro M, Saraiva AM, Imperatriz-Fonseca VL, Giannini TC (2018) Bat diversity in Carajás National Forest (Eastern Amazon) and potential impacts on ecosystem services under climate change. Biological Conservation 218: 200–210. https://doi.org/10.1016/j.biocon.2017.12.034

8. Gastauer M, Vero MPO, de Souza KP, Pires ES, Alves R, Caldeira CF, Ramos SJ, Oliveira G (2019) A metagenomic survey of soil microbial communities along a rehabilitation chronosequence after iron ore mining. Scientific Data 6:190008. https://doi.org/10.1038/sdata.2019.8

Written by

Guilherme Oliveira

Guilherme Oliveira

Environmental Genomics Group, Instituto Tecnológico Vale, Belém, Brazil

Gisele Nunes Lopes

Gisele Nunes Lopes

Environmental Genomics Group, Instituto Tecnológico Vale, Belém, Brazil

Rafael Valadares

Rafael Valadares

Environmental Genomics Group, Instituto Tecnológico Vale, Belém, Brazil

Ronnie Alves

Ronnie Alves

Environmental Genomics Group, Instituto Tecnológico Vale, Belém, Brazil

Santelmo Vasconcelos

Santelmo Vasconcelos

Environmental Genomics Group, Instituto Tecnológico Vale, Belém, Brazil

April 7, 2019
PDF
https://doi.org/10.21083/ibol.v9i1.5498

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