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      Is genetic engineering ever going to take off in forage, turf and bioenergy crop breeding?

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          Abstract

          Background

          Genetic engineering offers the opportunity to generate unique genetic variation that is either absent in the sexually compatible gene pool or has very low heritability. The generation of transgenic plants, coupled with breeding, has led to the production of widely used transgenic cultivars in several major cash crops, such as maize, soybean, cotton and canola. The process for regulatory approval of genetically engineered crops is slow and subject to extensive political interference. The situation in forage grasses and legumes is more complicated.

          Scope

          Most widely grown forage, turf and bioenergy species (e.g. tall fescue, perennial ryegrass, switchgrass, alfalfa, white clover) are highly self-incompatible and outcrossing. Compared with inbreeding species, they have a high potential to pass their genes to adjacent plants. A major biosafety concern in these species is pollen-mediated transgene flow. Because human consumption is indirect, risk assessment of transgenic forage, turf and bioenergy species has focused on their environmental or ecological impacts. Although significant progress has been made in genetic modification of these species, commercialization of transgenic cultivars is very limited because of the stringent and costly regulatory requirements. To date, the only transgenic forage crop deregulated in the US is ‘Roundup Ready’ (RR) alfalfa. The approval process for RR alfalfa was complicated, involving several rounds of regulation, deregulation and re-regulation. Nevertheless, commercialization of RR alfalfa is an important step forward in regulatory approval of a perennial outcrossing forage crop. As additional transgenic forage, turf and bioenergy crops are generated and tested, different strategies have been developed to meet regulatory requirements. Recent progress in risk assessment and deregulation of transgenic forage and turf species is summarized and discussed.

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          Most cited references102

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          Net energy of cellulosic ethanol from switchgrass.

          Perennial herbaceous plants such as switchgrass (Panicum virgatum L.) are being evaluated as cellulosic bioenergy crops. Two major concerns have been the net energy efficiency and economic feasibility of switchgrass and similar crops. All previous energy analyses have been based on data from research plots (<5 m2) and estimated inputs. We managed switchgrass as a biomass energy crop in field trials of 3-9 ha (1 ha = 10,000 m2) on marginal cropland on 10 farms across a wide precipitation and temperature gradient in the midcontinental U.S. to determine net energy and economic costs based on known farm inputs and harvested yields. In this report, we summarize the agricultural energy input costs, biomass yield, estimated ethanol output, greenhouse gas emissions, and net energy results. Annual biomass yields of established fields averaged 5.2-11.1 Mg x ha(-1) with a resulting average estimated net energy yield (NEY) of 60 GJ x ha(-1) x y(-1). Switchgrass produced 540% more renewable than nonrenewable energy consumed. Switchgrass monocultures managed for high yield produced 93% more biomass yield and an equivalent estimated NEY than previous estimates from human-made prairies that received low agricultural inputs. Estimated average greenhouse gas (GHG) emissions from cellulosic ethanol derived from switchgrass were 94% lower than estimated GHG from gasoline. This is a baseline study that represents the genetic material and agronomic technology available for switchgrass production in 2000 and 2001, when the fields were planted. Improved genetics and agronomics may further enhance energy sustainability and biofuel yield of switchgrass.
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            Overexpression of WXP1, a putative Medicago truncatula AP2 domain-containing transcription factor gene, increases cuticular wax accumulation and enhances drought tolerance in transgenic alfalfa (Medicago sativa).

            The identification of leaf wax genes involved in stress tolerance is expected to have great potential for crop improvement. Here we report the characterization of a novel AP2 domain-containing putative transcription factor gene from the model legume Medicago truncatula. The gene, designated WXP1, is able to activate wax production and confer drought tolerance in alfalfa (Medicago sativa), the most important forage legume species in the world and a close relative of M. truncatula. The predicted protein of WXP1 has 371 aa; it is one of the longest peptides of all the single AP2 domain proteins in M. truncatula. WXP1 is distinctly different from the most studied genes in the AP2/ERF transcription factor family such as AP2s, CBF/DREB1s, DREB2s, WIN1/SHN1 and GL15. Transcript level of WXP1 is inducible by cold, abscisic acid and drought treatment mainly in shoot tissues in M. truncatula. Overexpression of WXP1 under the control of the CaMV35S promoter led to a significant increase in cuticular wax loading on leaves of transgenic alfalfa. Scanning electron microscopy revealed earlier accumulation of wax crystals on the adaxial surface of newly expanded leaves and higher densities of wax crystalline structures on both adaxial and abaxial surfaces of mature leaves. Gas chromatography-mass spectrometry analysis revealed that total leaf wax accumulation per surface area increased 29.6-37.7% in the transgenic lines, and the increase was mainly contributed by C30 primary alcohol. WXP1 overexpression induced a number of wax-related genes. Transgenic leaves showed reduced water loss and chlorophyll leaching. Transgenic alfalfa plants with increased cuticular waxes showed enhanced drought tolerance demonstrated by delayed wilting after watering was ceased and quicker and better recovery when the dehydrated plants were re-watered.
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              Overexpression of miR156 in switchgrass (Panicum virgatum L.) results in various morphological alterations and leads to improved biomass production

              Switchgrass (Panicum virgatum L.) has been developed into a dedicated herbaceous bioenergy crop. Biomass yield is a major target trait for genetic improvement of switchgrass. microRNAs have emerged as a prominent class of gene regulatory factors that has the potential to improve complex traits such as biomass yield. A miR156b precursor was overexpressed in switchgrass. The effects of miR156 overexpression on SQUAMOSA PROMOTER BINDING PROTEIN LIKE (SPL) genes were revealed by microarray and quantitative RT-PCR analyses. Morphological alterations, biomass yield, saccharification efficiency and forage digestibility of the transgenic plants were characterized. miR156 controls apical dominance and floral transition in switchgrass by suppressing its target SPL genes. Relatively low levels of miR156 overexpression were sufficient to increase biomass yield while producing plants with normal flowering time. Moderate levels of miR156 led to improved biomass but the plants were non-flowering. These two groups of plants produced 58%–101% more biomass yield compared with the control. However, high miR156 levels resulted in severely stunted growth. The degree of morphological alterations of the transgenic switchgrass depends on miR156 level. Compared with floral transition, a lower miR156 level is required to disrupt apical dominance. The improvement in biomass yield was mainly because of the increase in tiller number. Targeted overexpression of miR156 also improved solubilized sugar yield and forage digestibility, and offered an effective approach for transgene containment.
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                Author and article information

                Journal
                Ann Bot
                Ann. Bot
                annbot
                annbot
                Annals of Botany
                Oxford University Press
                0305-7364
                1095-8290
                November 2012
                28 February 2012
                28 February 2012
                : 110
                : 6
                : 1317-1325
                Affiliations
                Forage Improvement Division, The Samuel Roberts Noble Foundation, Ardmore, OK 73401, USA
                Author notes
                [* ]For correspondence. Email: zywang@ 123456noble.org
                Article
                mcs027
                10.1093/aob/mcs027
                3478041
                22378838
                01d1bd91-21aa-4634-95ec-71c5cde7049a
                © The Author 2012. Published by Oxford University Press on behalf of the Annals of Botany Company.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 1 December 2011
                : 22 December 2011
                : 5 January 2012
                Categories
                Breeding Strategies for Forage and Grass Improvement

                Plant science & Botany
                bioenergy crops,creeping bentgrass,kentucky bluegrass,transgenesis,alfalfa,roundup ready,forage and turf,biosafety,transgenic plant,legume,grass,gene flow,deregulation

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