FeedBack (from this point onward, referred to as 'dB') is a custom chart editing tool which was designed to simplify the process of making custom note charts for the Guitar Hero games, and it can be used to make customs for Frets On Fire too.FeedBack is currently the easiest way to create customs for Frets On Fire.Its interface is designed to be simple and easy+fast to use, but I understand it can be a little confusing to first time users.
Simply run FeedBack.exe, there is no installation required.When you start the application you will see the help screen, this displays your current key-map configuration (which can be edited in the Config.ini file if it is inappropriate for you for whatever reason).Press [Esc] to exit the help screen, and you will see the main editor window.This is where you edit your chart, syncing to music, placing notes, etc. More about editing charts later.So if you have gotten this far and the software is running correctly you will want to get started.
Gh3 Pc Editing Tool V0.32
AllIntrameans is that there is no interframe encoding. In other words each frame is a stand-alone, not using fancy data compression called LGOP (Long Group of Pictures) where the amount of compression used depends on how information much changes between frames. AVCHD is such a codec, and while very good, and sometimes indistinguishable from an Intra codec, it is not a very good editing codec. Until recently it required very fast computers indeed to edit AVCHD without transcodeing, and therefore almost everyone doesthat as a matter of course. This takes time.
Fruit crops, consist of climacteric and non-climacteric fruits, are the major sources of nutrients and fiber for human diet. Since 2013, CRISPR/Cas (Clustered Regularly Interspersed Short Palindromic Repeats and CRISPR-Associated Protein) genome editing system has been widely employed in different plants, leading to unprecedented progress in the genetic improvement of many agronomically important fruit crops. Here, we summarize latest advancements in CRISPR/Cas genome editing of fruit crops, including efforts to decipher the mechanisms behind plant development and plant immunity, We also highlight the potential challenges and improvements in the application of genome editing tools to fruit crops, including optimizing the expression of CRISPR/Cas cassette, improving the delivery efficiency of CRISPR/Cas reagents, increasing the specificity of genome editing, and optimizing the transformation and regeneration system. In addition, we propose the perspectives on the application of genome editing in crop breeding especially in fruit crops and highlight the potential challenges. It is worth noting that efforts to manipulate fruit crops with genome editing systems are urgently needed for fruit crops breeding and demonstration.
Over the long history of crop domestication, four major plant breeding techniques have been developed and exploited: 1) conventional breeding by crossing and selection; 2) mutation-based plant breeding; 3) transgene-based plant breeding; and 4) the genome editing-based plant breeding (Hickey et al. 2019). The traditional breeding by hybridization and mutation-based breeding usually take decades and are labor-intensive (Chen et al. 2019). The transgenic plant breeding developed rapidly since the last century and emerged as one of the most promising ways to accumulate several elite traits in one variety, even though this technology aroused a lot of controversies soon after its birth, mainly due to the safety concerns and ethical issues (Prado et al. 2014).
Genome editing has been developed to obtain desired plant traits, as it could generate precise genome modification. Many systems have been developed to achieve genome editing in plants, including zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and the clustered regularly interspersed short palindromic repeats (CRISPR)/Cas system (Voytas and Gao 2014). CRISPR/Cas system has become one of the most widely used systems due to its low-cost, easy-to adapt and high specificity during genetic manipulation (Yin et al. 2017). This technology has been successfully applied in many cereals and economically important crops such as rice (Oryza sativa) (Shan et al. 2013), wheat (Triticum aestivum) (Shan et al. 2013), maize (Zea mays), potato (Solanum tuberosum) (Wang et al. 2015a), cassava (Manihot esculenta) (Odipio et al. 2017), chrysanthemums (Chrysanthemum morifolium) (Kishi-Kaboshi et al. 2017), European chestnut (Castanea sativa) (Pavese et al. 2021), Kabuli chickpea (Cicer arietinum) (Badhan et al. 2021), poinsettias (Euphorbia pulcherrima) (Nitarska et al. 2021) and rose (Rosa hybrida) (Wang et al. 2022a) (Fig. 1). Here we mainly describe status of CRISPR/Cas based genome editing in various fruit crops, emphasizing on plant development and disease resistance, especially plant architecture, fruit development, fruit ripening and quality, biotic stresses, and abiotic stresses in climacteric and non-climacteric fruits. We propose the future directions, especially the application of genome editing to promote plant breeding and overcome obstacles in fruit crops production.
Classical genome editing is accomplished via the DNA repair pathway after DNA double strand breaks (DSBs). Targeted DSBs are caused by sequence-specific nucleases (SSNs) enzymes which recognize and break DNA strand specifically (Voytas and Gao 2014). Four different SSNs have been adopted to introduce DSBs, including meganucleases, zinc finger nucleases (ZFNs) (Fig. 2A), transcription activator-like effector nucleases (TALENs) (Fig. 2B), and CRISPR/Cas reagents (Fig. 2C). When SSNs recognize and introduce DSBs, they are repaired by endogenous DNA repair pathways including non-homologous end joining (NHEJ) and homology-directed repair (HDR). The NHEJ repair pathway does not require a homologous repair template and usually introduces small insertions, deletions, or substitutions, eventually leading to genome modification and loss of gene function (Chen et al. 2019) (Fig. 2G). On the contrary, the HDR repair pathway requires a homologous DNA template to introduce insertions, mutations or replacements of DNA fragments (Gao 2021) (Fig. 2G). The application of meganucleases, ZFNs and TALENs is limited mainly due to the low specificity or efficiency when they recognize and cleave DNA targets through protein-DNA interactions. In contrast, the recently developed CRISPR/Cas system is more convenient and efficient in mediating genome modification in different plants.
Among different genome editing technologies, CRISPR/Cas9 system, and it derived base editor and prime editor systems have become powerful tools for gene editing. Here, we mainly focus on the application of CIRPSR/Cas system as well as base editor in the field of economically important fruit crops (Table 1). We mainly describe the status of the application of these technologies in plant development and plant resistance. Subsequently, we highlight future perspective on applying genome editing for fruit crop improvement, in order to provide insights for future plant breeding in fruit crops.
Fruit crops are major economic crops in many countries or regions in the world and are of great importance to the global economy and human nutrients. Fleshy fruits are divided into climacteric and non-climacteric types based on different respiratory features and ethylene biosynthesis rates during fruit ripening (Giovannoni 2001). In climacteric fruits such as apple, banana, tomato, kiwifruit and peach, the gaseous hormone ethylene is required for fruit ripening (Brumos 2021). Whereas in non-climacteric fruits such as strawberry, grape, watermelon, cucumber and citrus, fruit ripening is mainly regulated by abscisic acid (ABA), and ethylene is dispensable for the ripening process (Giovannoni 2001; Cherian et al. 2014). Many efforts have been made to optimize plant genome editing systems to improve fruit quality in climacteric fruits and non-climacteric fruits.
CRISPR/Cas9-mediated gene-edited fruits with desirable quality traits are in great demand. Cucumber has a special warty trait, consisting of spines and tubercules that greatly affect fruit appearance and market value. CsHEC2, a basic Helix-Loop-Helix (bHLH) gene regulating wart formation, was edited with CRISPR/Cas9. Two homozygous T2 cshec mutant lines exhibited reduced wart density (Wang et al. 2021e). A more detailed mechanism of wart development was investigated by editing genes such as Csa2G264590 (Liu et al. 2022). CRISPR/Cas9-mediated Csa2G264590 mutation led to an increase in spine number over ten times that of the wild type (Liu et al. 2022). These works demonstrated that the CRISPR/Cas9 system has advanced improvements in cucumber fruit quality and is of great value for cucumber breeding.
Watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai) is the second species in the Cucurbitaceae family with successful genome editing (Zhou et al. 2020b), and researchers have mainly focused on editing genes involved in watermelon flower sex determination. Genome editing of a C2H2 zinc finger transcription factor ClWIP1 led to the formation of gynoecious watermelon, with female and hermaphroditic flowers observed in homozygous T1 mutants (Zhang et al. 2020a).
The CRISPR/Cas9 system has also been applied for watermelon fruit quality improvement. Sugar content is mediated by NAC (NAM, ATAF1/2, and CUC2) transcription factors. CRISPR/Cas-mediated genome editing of the ClNAC68 resulted in decreased contents of fructose, glucose, and sucrose accumulation (Wang et al. 2021b). Moreover, Melatonin (N-acetyl-5-methoxytryptamine) is identified as a key bioactive molecule involved in many processes in plants and animals (Zhang et al. 2015b). Caffeic acid O-methyltransferase gene ClCOMT1 plays a key role in melatonin biosynthesis (Kang et al. 2013b). CRISPR/Cas9 mediated clcomt1 mutation led to decreased melatonin contents in watermelon calli (Chang et al. 2021). In addition, the CRISPR/Cas system has been used to target the ABA hydrolyzation β-glucosidase (BG) gene (Wang et al. 2021c) to generate transgene-free bg1 mutants with reduced seed size and weight. 2ff7e9595c
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